The DOI matches its Crossref record; 77 of the 77 references it cites were found in a registry or an index; 53 of the 110 sources its claims cite were checked for support.
Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids
Open accessAbstract
Recombinant adeno-associated virus vectors (rAAV) are being explored as gene delivery vehicles for the treatment of various inherited and acquired disorders. rAAVs are attractive vectors for several reasons: wild-type AAVs are nonpathogenic, and rAAVs can trigger long-term transgene expression even in the absence of genome integration-at least in postmitotic tissues. Moreover, rAAVs have a low immunogenic profile, and the various AAV serotypes and variants display broad but distinct tropisms. One limitation of rAAVs is that their genome-packaging capacity is only ∼5 kb. For most applications this is not of major concern because the median human protein size is 375 amino acids. Excluding the ITRs, for a protein of typical length, this allows the incorporation of ∼3.5 kb of DNA for the promoter, polyadenylation sequence, and other regulatory elements into a single AAV vector. Nonetheless, for certain diseases the packaging limit of AAV does not allow the delivery of a full-length therapeutic protein by a single AAV vector. Hence, approaches to overcome this limitation have become an important area of research for AAV gene therapy. Among the most promising approaches to overcome the limitation imposed by the packaging capacity of AAV is the use of dual-vector approaches, whereby a transgene is split across two separate AAV vectors. Coinfection of a cell with these two rAAVs will then-through a variety of mechanisms-result in the transcription of an assembled mRNA that could not be encoded by a single AAV vector because of the DNA packaging limits of AAV. The main purpose of this review is to assess the current literature with respect to dual-AAV-vector design, to highlight the effectiveness of the different methodologies and to briefly discuss future areas of research to improve the efficiency of dual-AAV-vector transduction.
Overview
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Written from the paper’s full text.
TL;DR
This review assesses strategies for expressing transgenes that exceed the ~5 kb packaging capacity of recombinant AAV vectors, focusing on dual-vector designs (fragmented/oversized, overlapping, trans-splicing, and hybrid). It concludes that hybrid dual-vector systems are currently the most promising in most instances, but that no single system is optimal across all transgenes and tissues.
Key findings
- AAV packaging capacity is limited to roughly 5 kb: wtAAV genomes average ~4.7 kb, Hermonat et al. found ~5.6 kb as the maximum for recovery of significant infectious virus, and Dong et al. reported optimal packaging between 4.1 and 4.9 kb, with vectors >5.2 kb packaged very inefficiently. Claims that AAV5 could package up to 8.9 kb were not supported by later studies, which found such genomes truncated (packaged genomes never exceeded 5.2 kb).
- Because ~6% of human proteins have coding sequences exceeding 4 kb (median human protein is 375 amino acids), some therapeutic cDNAs (e.g., ABCA4 ~7.3 kb, dystrophin, dysferlin, CFTR, MYO7A, factor VIII) cannot fit in a single AAV vector.
- In a direct comparison (Ghosh et al.), hybrid vectors achieved LacZ expression equal to a single intact vector at MOI 20,000 vg/cell, and 71% at 1,000 vg/cell, outperforming overlapping (~25%) and trans-splicing (~50%) variants; in mouse tibialis anterior muscle, hybrid vectors yielded 81% of single-vector β-galactosidase activity versus 34% (overlapping) and 62% (trans-splicing).
- Hybrid systems are not universally superior: Zhang and Duan found a dual overlapping vector system gave ~3-fold higher muscle transduction than three hybrid systems, and Pryadkina et al. obtained similar results for dysferlin. Minimized AP bridging sequences (0.27/0.26 kb vs. 0.87 kb) expand payload by ~1.2 kb; a 77 bp F1 phage-derived homology region has also been used.
Methods
- Narrative literature review; no primary experimental data reported.
- Synthesizes published studies on AAV biology, packaging capacity limits, and dual/triple-vector strategies (fragmented/oversized, overlapping, trans-splicing, hybrid), including in vitro (e.g., HEK293, MO59K, C2C12) and in vivo (mouse models including mdx, mdx4cv, A/J, ABCA4−/−, sh1) comparisons.
- Discusses mechanistic hypotheses (homologous recombination, ITR-mediated concatamerization, Rad51C-dependent repair) and design variables (recombinogenic sequences, splice donor/acceptor sites, ITR heterology).
Limitations
- Stated by the paper: the literature on dual-vector efficacy is "broad and often conflicting"; the mechanism of transduction by fragmented and overlapping vectors is "as yet not fully understood"; efficiency of transgene expression from dual-vector systems has been inconsistent and is currently insufficient for many clinical applications; hybrid systems are not always the most efficient; and optimization must be done empirically per transgene and tissue.
- Evident from the text: as a narrative review it provides no new experimental data, and several cited comparisons rely on different transgenes, serotypes, cell/tissue types, and titration methods, which the authors note can themselves influence results.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
53 of 110 cited sources checked · 34 supported · 0 flagged · accuracy not yet measured
Every source the paper cites for this claim was checked and supports it.
In the paper
“Trapani et al. found the overexpression of MYO7a in HEK293 cells with AAV2 vectors to be more efficient when they used a dual overlapping vector system as opposed to a single oversized vector, 46”
What the sources say
- Supportedcites [46] Trapani I, 2014, EMBO Mol Med source
The Results section, “Dual AAV vectors allow high levels of transduction in vitro,” reports that HEK293 cells were infected with AAV2/2 vectors encoding MYO7A. Its quantification of MYO7A expression found the dual overlapping (OV) approach more efficient than the single oversized (OZ) approach. (LLM, conf=0.94)
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passes- Supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“even such seemingly arcane technical details as the method of vector titration appear to be able to influence the results obtained when comparing two different strategies to increase AAV payload. 46”
What the sources say
- Could not verifycites [46] Trapani I, 2014, EMBO Mol Med sourceExperimental — accuracy not yet measured
The Materials and Methods section describes how vector titers were measured, and the Results compare strategies for delivering large genes. Neither passage says that the method of titration influences the comparison. The Discussion suggests other possible reasons for differences between studies, including vector design and purification, but does not identify titration method as one. (LLM, conf=0.76)
Strength:hedged· not yet calibratedCausal:conditional· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passes- Could not verify
Every source the paper cites for this claim was checked and supports it.
In the paper
“Transduction of airway epithelial cells with an rAAV vector containing this expression cassette was sufficient to reverse, at least partially, the cystic fibrosis chloride transport defect in these cells. 35”
What the sources say
- Supportedcites [35] source
The Results and Discussion report that AAV5 vectors carrying the shortened CFTR expression cassette generated chloride current in differentiated primary CF airway epithelia, with correction persisting for at least 28 days. The same section concludes that a single AAV vector corrected the CF chloride-transport defect; the claim’s qualification, ‘at least partially,’ is consistent with this result. (LLM, conf=0.94)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passes- Supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“the median length of a human protein is only 375 amino acids. 27”
What the sources say
- Could not verifycites [27] sourceExperimental — accuracy not yet measured
The Results section (‘Protein lengths across proteomes’; page unavailable) reports a median of 361 amino acids for the pooled eukaryotic proteins, not a human-specific median. The Methods section (‘Proteomes’; page unavailable) says human proteins were included, but searches for ‘375’ and ‘Homo sapiens Table 2 median’ found no passage in our copy giving the claimed human value. (LLM, conf=0.72)
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passes- Could not verify
Every source the paper cites for this claim was checked and supports it.
In the paper
“Hu and Lipshutz used a similar dual-vector strategy to demonstrate phenotypic correction of FVIII deficiency in a mouse model of hemophilia A. 39”
What the sources say
- Supportedcites [39] source
The Introduction describes co-injecting two AAV vectors, one expressing the FVIII heavy chain and the other the light chain, in a murine model of hemophilia A. The Results report improved FVIII activity and shortened bleeding time in treated knockout mice, demonstrating partial phenotypic correction. (LLM, conf=0.94)
Strength:boosted· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passes- Supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“in mdx mice (a mouse model of Duchenne muscular dystrophy) gene transfer with AAV vectors encoding microdystrophin provides significant, although not optimal therapeutic benefit. 40-42”
What the sources say
- Supportedcites [40] source
The Results (page not supplied) report significant improvements in cardiac function after AAV-microdystrophin treatment of aged female mdx mice. They also state that some measures did not return to wild-type levels, supporting the qualification that the benefit was not optimal. (LLM, conf=0.91)
- Not checked — no full text of the sourcecites [41] source
- Not checked — no full text of the sourcecites [42] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passes- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“During transduction, rAAV genomes have been shown to form circular or linear (mostly) head-to-tail arrays of viral genomes, 14,50,51”
What the sources say
- Partly supportedcites [14] Duan D, 1998, J Virol sourceExperimental — accuracy not yet measured
The Results and Discussion report head-to-tail circular rAAV concatemers in infected muscle tissue. The Results say that linear integrated concatemers had previously been hypothesized, whereas this study's data suggest the persistent genomes are circular and episomal. The claim's assertion that linear arrays have been shown to form is therefore not established by this paper. (LLM, conf=0.86)
- Partly supportedcites [50] sourceExperimental — accuracy not yet measured
The Abstract reports circular rAAV intermediates after transduction and describes monomer and multimer head-to-tail conformations. The paper does not establish that linear arrays formed during transduction are mostly head-to-tail; its statement about the most abundant head-to-tail form is limited to a model of circular genomes. (LLM, conf=0.85)
- Not checked — no full text of the sourcecites [51] source
Load-bearingStrength:boosted· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passes- Partly supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“it has been reported that the CMV promoter can be shortened to 173 bp without apparent loss in expression levels-at least in certain cell types. 35”
What the sources say
- Supportedcites [35] source
The Results and Discussion report that the 173CMV promoter produced β-gal activity equivalent to the full-length CMVie enhancer/promoter in A549 cells and primary airway epithelial cultures. It produced less activity in H441 cells, consistent with the claim’s qualification that expression was maintained only in certain cell types. (LLM, conf=0.94)
Load-bearingStrength:hedged· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passes- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“AAV vectors with two complementary ITRs can form circular concatamers. 13-16”
What the sources say
- Not checked — no full text of the sourcecites [13] source
- Supportedcites [14] Duan D, 1998, J Virol source
The Results describe recombinant AAV circular intermediates with two ITRs in a head-to-tail arrangement and identify larger circular multimers containing approximately three AAV genomes. This supports the claim that AAV vectors with ITRs can form circular concatemers. Page numbers are unavailable in our copy. (LLM, conf=0.88)
- Not checked — no full text of the sourcecites [15] source
- Partly supportedcites [16] sourceExperimental — accuracy not yet measured
The Discussion (page unavailable) says recombinant AAV derivatives can assume circular multimeric structures, supporting the circular-concatamer part of the claim. It also suggests that ITRs contribute to circle formation, but does not establish the specific condition of two complementary ITRs. The concatameric arrays described in the Results were linear products of rolling-circle amplification, not circular vector concatamers. (LLM, conf=0.83)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesThe cited sources support only part of this claim — read each source’s verdict below.
In the paper
“overexpression of dysferlin by intramuscular injection of AAV8 encoding dysferlin from either a single oversized vector or from two overlapping vectors showed a * 10-fold higher expression of dysferlin with the dual-vector system. 45”
What the sources say
- Partly supportedcites [45] sourceExperimental — accuracy not yet measured
The Results section on intramuscular injection reports higher dysferlin expression with the overlapping dual-vector Set 3 than with the oversized fAAV vector. It gives the increase as approximately 4–10-fold relative to Batch 2 fAAV, so the citing sentence’s approximately 10-fold figure omits the lower end of the reported range. (LLM, conf=0.87)
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passes- Partly supported
- It may state two separate claims as one. 26
- Probably not a substantial claim. 8
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“it is generally believed that rAAV genomes persist episomally as mostly circular concatamers 16 in postmitotic cells, leading to long-term transgene expression.”
What the sources say
- Partly supportedcites [16] sourceExperimental — accuracy not yet measured
The Discussion (page unavailable) reports that naturally acquired wild-type AAV persists mainly as circular episomes in human tissues and says this is consistent with circular episomal forms of recombinant AAV vectors. It does not establish that recombinant genomes are mostly circular concatamers in postmitotic cells or that this form leads to long-term transgene expression. The concatameric forms described in the Results are products of rolling-circle amplification, not evidence that the persisting genomes have that structure. (LLM, conf=0.86)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
- Partly supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“However, the finding that AAV5 capsids could package intact genomes of up to 8.9 kb in length was not supported by further studies, which found that such genomes were truncated and that the packaging capacity of AAV was still limited to genomes substantially smaller than 8.9 kb. 30-32”
What the sources say
- Not checked — no full text of the sourcecites [30] source
- Supportedcites [31] source
Split into 2 atoms — 2 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
- Supportedcites [32] source
Split into 2 atoms — 2 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
Every source the paper cites for this claim was checked and supports it.
In the paper
“Samulski and colleagues reported that transduction by oversized vectors depends on the activity of the strand-annealing protein Rad51C but is independent of the catalytic subunit of DNA-dependent protein kinase. 34”
What the sources say
- Supportedcites [34] source
The Results (p. 2210) report a 50-fold decrease in fragment AAV (fAAV) transduction without Rad51C, while loss of DNA-PKcs did not significantly alter it. The paper distinguishes these oversized-vector fragments from split AAV vectors, which do depend on DNA-PKcs. (LLM, conf=0.93)
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✓ Based: passesProbably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“similar to dual overlapping vectors, trans-splicing vector-pair systems do not always outperform single oversized vectors. For instance, Hirsch et al. 34 reported higher expression both in vitro (C2C12 myoblasts and myotubules) and in vivo with a single oversized genome vector when compared with a transsplicing system. 34”
What the sources say
- Supportedcites [34] source
In “Comparison of AAV large-gene transduction strategies” (pp. 2209–2210), the paper reports significantly greater transduction with the oversized fragment AAV vector than with split (trans-splicing) AAV in C2C12 myoblasts and myotubules. In vivo, it reports fourfold higher luciferase activity in mouse muscle at the lower dose and sevenfold greater transduction efficiency in the retina. Muscle activity was similar at the higher dose, consistent with the claim’s qualification that trans-splicing systems do not always outperform oversized vectors. (LLM, conf=0.93)
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passesIt may state two separate claims as one.
- Supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“The trans-splicing method to expand the size of ORFs that can be successfully expressed using AAV vectors was originally developed in the laboratory of John Engelhardt 52,53 and has since been utilized in several studies with varying success (e.g., refs. 53-57 ).”
What the sources say
- Not checked — no full text of the sourcecites [52] source
- Supportedcites [53] source
The Abstract (page not provided) says the authors developed a two-vector trans-splicing approach to deliver transgenes beyond AAV’s packaging limit and demonstrated functional expression. John Engelhardt is among the paper’s authors. The sentence cites other studies for its separate claim about subsequent uses. (LLM, conf=0.90)
- Not checked — no full text of the sourcecites [54] source
- Not checked — no full text of the sourcecites [55] source
- Not checked — no full text of the sourcecites [56] source
- Not checked — no full text of the sourcecites [57] source
Strength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✓ Based: passesProbably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
Every source the paper cites for this claim was checked and supports it.
In the paper
“Consistent with their in vitro results, dual trans-splicing and hybrid-AK vectors successfully transduced mouse and pig photoreceptors and were found to improve the phenotype of mouse models of STGD and USH1B. 46”
What the sources say
- Supportedcites [46] Trapani I, 2014, EMBO Mol Med source
The Results section, “Dual AAV TS and hybrid AK but not OV vectors transduce mouse and pig photoreceptors” (page unavailable), reports photoreceptor transduction in both species. The Results section, “Dual AAV vectors improve the retinal phenotype of STGD and USH1B mouse models” (page unavailable), reports improved retinal measures after either vector treatment in the STGD and USH1B models. (LLM, conf=0.94)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“Interestingly, however, both Dyka et al. 47 and Trapani et al. 46 observed the expression of both full-length and smaller fragments (of defined size) of MYO7A and or ABCA4 with the hybrid dual-vector system.”
What the sources say
- Supportedcites [47] source
The Results section (‘Comparison of relative efficiencies and specificity of full-length MYO7A expression,’ Fig. 4; page not supplied) reports full-length MYO7A from the AP hybrid platform and a defined, less-than-full-length peptide detectable on Western blot. This confirms the observation attributed to Dyka et al.; the claim about Trapani et al. is not assessed against this source. (LLM, conf=0.92)
- Supportedcites [46] Trapani I, 2014, EMBO Mol Med source
The Results ('Dual AAV vectors allow high levels of transduction in vitro'; page unavailable) report full-length ABCA4 and MYO7A proteins from hybrid dual vectors. The Results on the mouse models and the Discussion (pages unavailable) also report smaller MYO7A proteins from vector halves in vitro, while specifying that only full-length proteins were detected in the retina in vivo. (LLM, conf=0.90)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
- Supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“Because such a circularization is an intramolecular event, it is probably more efficient than intermolecular concatemerization. 34,49,58”
What the sources say
- Supportedcites [34] source
The Introduction (p. 2205) describes circularization as intramolecular, rapid, and initially the primary persistent form of rAAV DNA, while intermolecular linkages occur to a lesser extent. It also says AAV genomes preferentially form circular monomers, supporting the claim’s qualified comparison. (LLM, conf=0.90)
- Not checked — no full text of the sourcecites [49] source
- Not checked — no full text of the sourcecites [58] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“But they also demonstrate that there is no one-fits-all dual-vector system, and that the nature of the artificial recombinogenic regions and the tissue being targeted can drastically influence the efficiency of transduction with such hybrid split AAV vector systems. 46”
What the sources say
- Supportedcites [46] Trapani I, 2014, EMBO Mol Med source
The Results (Dual AAV vectors allow high levels of transduction in vitro; page unavailable) report that hybrid AP produced the lowest transgene-expression levels, while hybrid AK was more efficient. The Discussion (page unavailable) reports that hybrid AK outperformed trans-splicing in mouse photoreceptors but not retinal pigment epithelium, and suggests that differences in AP sequences may explain differing transduction levels across studies. Together, these passages support the claim that recombinogenic-region design and target tissue influence efficiency. (LLM, conf=0.90)
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
- Supported
In the paper
“Nonetheless, AAV5vectorswith the supposedly 8.9kb large viral genomes clearly did result in expression of ABCA4, MYO7A, and CEP290 from genomes whose intact genome length would be 8.9, 8.1, and 8.9 kb in size, respectively.”
What the sources say
- Not checked — no full text of the sourcecites [29] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“the efficiency of reconstituting a genome with a full-length expression cassette is likely dependent on the number of nuclear copies of vector genomes.”
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✗ Based: failsProbably not a substantial claim. The paper may give no reason why it holds.
In the paper
“vectors larger than 5.2 kb were found to be packaged very inefficiently, and transduction efficiencies of these oversized vectors were dramatically reduced. 26”
What the sources say
- Not checked — no full text of the sourcecites [26] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“Hermonat et al. concluded that the maximum genome size that allowed the recovery of significant amounts of infectious virus was approximately 5.6 kb. 25”
What the sources say
- Not checked — no full text of the sourcecites [25] source
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesProbably not a substantial claim.
In the paper
“Southern blot hybridization revealed that purportedly oversized packaged genomes were in fact truncated, and the authors suggested that the truncated fragments could assemble thus restoring the full-length expression cassette.”
What the sources say
- Not checked — no full text of the sourcecites [30] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesProbably not a substantial claim. It may state two separate claims as one.
In the paper
“both AP expression and LacZ expression were detected in mdx muscle of adult mice injected with such AAV vector preparations.”
What the sources say
- Not checked — no full text of the sourcecites [30] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“the combination of AAV serotype and tissue/cell type might have a particularly profound effect on the efficiency of transduction with overlapping systems because for dual -vector approaches to be successful, a high level of co-transduction-preferably with multiple copies of each vector-is essential for efficient transgene overexpression.”
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“Although the overall levels of minidystrophin expression were lower than in wildtype muscles, there was an improvement in limb muscle physiological performance. 42”
What the sources say
- Not checked — no full text of the sourcecites [42] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“They could demonstrate improved expression with vectors containing this artificial intron when compared with trans-splicing vectors that contained a poor splice site; however, expression was not as efficient as with the best trans-splicing vectors using natural dystrophin exon/intron sites. 62”
What the sources say
- Not checked — no full text of the sourcecites [62] source
Strength:hedged· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passesIt may state two separate claims as one.
In the paper
“As expected, successful trans-splicing resulted in the expression of full-length dysferlin, leading to improved membrane repair capacity, muscle histology, and muscle function. 63”
What the sources say
- Not checked — no full text of the sourcecites [63] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“Expression from the single vector with a ''regular size genome'' and the hybrid vectors was strong and widespread, whereas expression from the transsplicing, and especially the overlapping vectors, was not as effective.”
What the sources say
- Not checked — no full text of the sourcecites [65] source
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“Although full-length dystrophin could be detected with this three-vector systemboth in vitro and in vivo -the expression was very weak.”
What the sources say
- Not checked — no full text of the sourcecites [70] source
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Maybe the firmest conclusion that can be drawn from the vast trove of available literature is that there is no one ideal system, and that the choice of what the best vector system is can vary drastically depending on the transgene and the cell type/ tissue targeted by the AAV vectors.”
Load-bearingStrength:hedged· not yet calibratedTests✓ Asserted: passes✗ Checkable: fails✓ Load-bearing: passes✓ Based: passesProbably not a substantial claim. It may be too vague to check against evidence.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“{ Strictly speaking, trans-splicing and hybrid vector systems do not rely on trans-splicing of pre-mRNAs. Instead, in both systems a full-length mRNA is generated whose manufactured intron is then removed by regular cis -splicing to join the two exons.”
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✗ Based: failsProbably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds. It may state two separate claims as one.
In the paper
“When b -galactosidase activity from muscle tissue was quantified, the hybrid vectors yielded 81% of the activity of the single ''regular size genome'' vector, which was significantly greater than the b -galactosidase activity from overlapping (34%) or trans-splicing vectors (62%).”
What the sources say
- Not checked — no full text of the sourcecites [65] source
Load-bearingStrength:plain· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“Either pathway results in LacZ expression.”
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes? Load-bearing: unclear✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Not only are such fragments likely nonfunctional, but they also could be dominant-negative and/or toxic.”
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes? Based: unclearIt may state two separate claims as one.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“The two vectors of this hybrid system can lead to the transcription of full-length mRNA by one of two ways: either through AP -mediated homologous recombination or via ITR-mediated head-to-tail concatamerization (Fig. 2).”
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
In the paper
“Thus, the hybrid vector outperformed both the overlapping and trans-splicing variants, which only expressed levels of approximately 25% and 50%, respectively, of the original construct.”
What the sources say
- Not checked — no full text of the sourcecites [65] source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In this context, the most important components of a dual hybrid vector system to be taken into account appear to be the choice of the recombinogenic region and the nature of splice donor and acceptor signals.”
Load-bearingStrength:hedged· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✗ Based: failsProbably not a substantial claim. The paper may give no reason why it holds. It may state two separate claims as one.
Citations
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“For many diseases, vector-mediated gene delivery is not feasible due to the cargo limit of viral vectors as well as the expression of multiple heterogeneous variants of the target gene [97,98].”
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“Full-length assembly can occur through recombination of homologous regions or/and trans-splicing (Chamberlain et al., 2016).”
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Adeno-Associated Virus Gene Therapy for Liver Disease
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“To overcome the size limitation challenge, dual vector systems have been proposed, where the larger genome would be recreated through recombination of the partial genomes.(32) In addition, the effort to develop cassettes for factor VIII (FVIII) expression serves as an object lesson in setting expectations for both eff…”
BackgroundViral Vectors for the in Vivo Delivery of CRISPR Components: Advances and Challenges
Mengstie, M. A. (2022) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2022.895713 · Cited by 146
“When both AAVs are used to transduce a cell, the full-length gene and/or protein are reconstituted (Chamberlain et al., 2016).”
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Ryanodine Receptor 1-Related Myopathies: Diagnostic and Therapeutic Approaches
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“RYR1 exceeds 159 kb and thus far supersedes the current packaging capacity for adenovirusmediated therapy (∼ 5 kb) [96].”
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“72 AAV-delivered transgenes form stable, nonreplicating circular episomal DNA, which is gradually lost in dividing cells, but persists in postmitotic cells, such as neurons, due to the very low cellular turnover.”
InfluentialCRISPR-Mediated Integration of Large Gene Cassettes Using AAV Donor Vectors
Bak, R. O., & Porteus, M. H. (2017) · Cell Reports · doi:10.1016/j.celrep.2017.06.064 · Cited by 124
“…recombinant AAV (rAAV) vectors has not been successfully extended, though strategies that enable episomal expression of transgenes that exceed the packaging capacity have been devised (Grieger and Samulski, 2005, Chamberlain et al., 2016, Nakai et al., 2000, Sun et al., 2000, Halbert et al., 2002).”
BackgroundAdeno-Associated Viral Vectors in Neuroscience Research
Haggerty, D. L., Grecco, G. G., Reeves, K. C., & Atwood, B. K. (2019) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2019.11.012 · Cited by 135
“Although AAV2 is the most widely used variant because it was the first serotype identified, numerous other serotypes with varying tropism exist, giving AAVs wide clinical applicability.(92,93,97) When combined, these characteristics have established AAVs as a leading vector choice for gene therapy studies with one of…”
BackgroundInfluentialChallenges for Therapeutic Applications of Opsin-Based Optogenetic Tools in Humans
Shen, Y., Campbell, R. E., Côté, D. C., & Paquet, M.-È. (2020) · Frontiers in Neural Circuits · doi:10.3389/fncir.2020.00041 · Cited by 119
“Existing serotypes have been ‘‘overpackaged’’ with mixed success and varying reproducibility and the consensus appears to be that AAV can be overpackaged by ∼10%, but with a concomitant reduction in both viral titers and in vivo transduction (Chamberlain et al., 2016).”
BackgroundSystemic delivery of adeno-associated viral vectors
Duan, D. (2016) · Current Opinion in Virology · doi:10.1016/j.coviro.2016.07.006 · Cited by 105
Current knowledge of SLC6A1-related neurodevelopmental disorders
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“, 2014) and (iii) the transporters’ relatively small size (599 aa) makes SLC6A1 a plausible candidate for viral-mediated gene therapy using adeno-associated virus vectors (Chamberlain et al., 2016).”
BackgroundAdeno-associated Virus (AAV) Dual Vector Strategies for Gene Therapy Encoding Large Transgenes.
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“This approach is carried out by a process called intein-mediated trans-splicing, which to some degree is similar to mRNA splicing [83, 84].”
BackgroundAn AAV Dual Vector Strategy Ameliorates the Stargardt Phenotype in Adult Abca4 −/− Mice
McClements, M. E., Barnard, A. R., Singh, M. S., Charbel Issa, P., Jiang, Z., Radu, R. A., & MacLaren, R. E. (2019) · Human Gene Therapy · doi:10.1089/hum.2018.156 · Cited by 92
Evaluating Efficiencies of Dual AAV Approaches for Retinal Targeting
Carvalho, L. S., Turunen, H. T., Wassmer, S. J., Luna-Velez, M. V., Xiao, R., Bennett, J., & Vandenberghe, L. H. (2017) · Frontiers in Neuroscience · doi:10.3389/fnins.2017.00503 · Cited by 86
“Consequently, there is great interest in increasing transgene size while still utilizing the AAV system and the use of a dual AAV vectors system has been extensively studied for different experimental and disease modalities (Chamberlain et al., 2016).”
BackgroundBarnes, L. F., Draper, B. E., Chen, Y., Powers, T. W., & Jarrold, M. F. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.08.002 · Cited by 87
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Cardiac gene therapy with adeno-associated virus-based vectors
Chamberlain, K., Riyad, J. M., & Weber, T. (2017) · Current Opinion in Cardiology · doi:10.1097/hco.0000000000000386 · Cited by 75
“On the contrary, for approximately onethird of the genes listed in Table 1, the size of the respective cDNA exceeds the packaging capacity of AAV, which is 5 kb (including ITRs and regulatory elements [11]).”
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“, protein trans-splicing) [38,60] have been pursued; however, these still occur with lowered efficiency and a clear upper packaging threshold at 5.”
BackgroundDirected Evolution of CRISPR/Cas Systems for Precise Gene Editing
Liu, R., Liang, L., Freed, E. F., & Gill, R. T. (2020) · Trends in biotechnology · doi:10.1016/j.tibtech.2020.07.005 · Cited by 58
Targeting Cancer with CRISPR/Cas9-Based Therapy
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Preclinical evaluation of the efficacy and safety of AAV1-hOTOF in mice and nonhuman primates
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“37 In the TS strategy, the fragment from splice donor (SD) to splice acceptor (SA) is spliced out by intracellular spliceosome after recombination and transcription of transgene, which generates a mature mRNA and translates functional protein.”
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The potential for targeted rewriting of epigenetic marks in COPD as a new therapeutic approach
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Dual and triple AAV delivery of large therapeutic gene sequences into the inner ear
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Development of a dual hybrid AAV vector for endothelial-targeted expression of von Willebrand factor
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Genetic Delivery and Gene Therapy in Pulmonary Hypertension
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Adeno-Associated Virus-Based Gene Therapy for Lifelong Correction of Genetic Disease
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“There are multiple strategies to circumvent the packaging limitation of AAV-based vector for delivery of Cas9 expression vectors (reviewed in Ref.(15)).”
BackgroundInfluentialAtaluren for the Treatment of Usher Syndrome 2A Caused by Nonsense Mutations
Samanta, A., Štingl, K., Kohl, S., Ries, J., Linnert, J., & Nagel‐Wolfrum, K. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20246274 · Cited by 39
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Dual-AAV delivery of large gene sequences to the inner ear
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Bankoti, K., Generotti, C. J., Hwa, T. P., Wang, L., O’Malley, B. W., & Li, D. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.03.005 · Cited by 42
Peptide ligands for the affinity purification of adeno‐associated viruses from HEK 293 cell lysates
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“AAV is a small, nonenveloped icosahedral virus, whose capsid can pack a linear single‐strand DNA (ssDNA) genome of up to about 5 kilobases (Chamberlain et al., 2016; Lai et al., 2010; Linden et al., 1996; Vasileva & Jessberger, 2005).”
BackgroundProtein Engineering Strategies to Expand CRISPR-Cas9 Applications
Ribeiro, L. F., Ribeiro, L. F. C., Barreto, M. Q., & Ward, R. J. (2018) · International Journal of Genomics · doi:10.1155/2018/1652567 · Cited by 36
“3 kb), which limits its efficient gene-based delivery via recombinant adenoassociated virus (rAAV) [80, 81].”
BackgroundGene delivery to the lungs: pulmonary gene therapy for cystic fibrosis
Villate-Beitia, I., Sesma, J. Z., Puras, G., & Pedraz, J. L. (2017) · Drug Development and Industrial Pharmacy · doi:10.1080/03639045.2017.1298122 · Cited by 38
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Disruptive Technology: CRISPR/Cas-Based Tools and Approaches
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Large gene delivery to the retina with AAV vectors: are we there yet?
Trapani, I., Tornabene, P., & Auricchio, A. (2020) · Gene Therapy · doi:10.1038/s41434-020-0174-4 · Cited by 30
Gene Transfer Therapy for Neurodevelopmental Disorders
Özlü, C., Bailey, R. M., Sinnett, S. E., & Goodspeed, K. (2021) · Developmental Neuroscience · doi:10.1159/000515434 · Cited by 31
Superior Fidelity and Distinct Editing Outcomes of SaCas9 Compared with SpCas9 in Genome Editing
Yang, Z.-X., Fu, Y.-W., Zhao, J.-J., Zhang, F., Li, S.-A., Zhao, M., Wen, W., Zhang, L., Cheng, T., Zhang, J., et al. (2022) · Genomics Proteomics & Bioinformatics · doi:10.1016/j.gpb.2022.12.003 · Cited by 32
“However, the large size of SpCas9 (1368 amino acids, (cid:1) 4.1 kb) limits its further applications for adeno-associated virus (AAV)- based in vivo gene therapy, as the limited cargo size of the AAV vector prevents co-packaging of both SpCas9 and sgRNA expression cassettes [10].”
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Fakhiri, J., & Grimm, D. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.04.005 · Cited by 29
Post-Vitrectomy Endophthalmitis in Victoria, Australia
Chandra, A., Smith, J. D., Wang, B. Z., Kurniawan, E. D., Kam, J., Sandhu, S. S., & Allen, P. J. (2016) · Asia-Pacific Journal of Ophthalmology · doi:10.22608/apo.2016126 · Cited by 28
Splicing in the pathogenesis, diagnosis and treatment of ciliopathies
Wheway, G., Lord, J., & Baralle, D. (2019) · Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms · doi:10.1016/j.bbagrm.2019.194433 · Cited by 27
Clinical and Translational Landscape of Viral Gene Therapies
Yudaeva, A., Kostyusheva, A., Kachanov, A., Brezgin, S., Ponomareva, N., Parodi, A., Pokrovsky, V. S., Lukashev, A. N., Chulanov, V., & Kostyushev, D. S. (2024) · Cells · doi:10.3390/cells13221916 · Cited by 30
Farzanehpour, M., Miri, A., Alvanegh, A. G., & Gouvarchinghaleh, H. E. (2023) · Biochemical Pharmacology · doi:10.1016/j.bcp.2023.115555 · Cited by 27
Canine Models of Inherited Musculoskeletal and Neurodegenerative Diseases
Story, B. D., Miller, M. E., Bradbury, A. M., Million, E. D., Duan, D., Taghian, T., Faissler, D., Fernau, D., Beecy, S. J., & Gray‐Edwards, H. L. (2020) · Frontiers in Veterinary Science · doi:10.3389/fvets.2020.00080 · Cited by 30
“A highly truncated, micro-dystrophin gene (<4 kb) (197, 198) or dual-AAV vectors used in combination to deliver a larger payload (199, 200).”
BackgroundThird-generation lentiviral gene therapy rescues function in a mouse model of Usher 1B
Schott, J. W., Huang, P., Morgan, M. A., Nelson‐Brantley, J., Koehler, A., Renslo, B., Büning, H., Warnecke, A., Schambach, A., & Staecker, H. (2023) · Molecular Therapy · doi:10.1016/j.ymthe.2023.10.018 · Cited by 25
“Although dual AAV vector systems, in which the transferred gene is split into two separate vectors that recombine in the target cell, 15 can allow the use of AAV vectors to target disorders with larger genes, this strategy may be associated with a loss in ef fi ciency.”
Dasari, B., Cashman, S. M., & Kumar‐Singh, R. (2017) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2017.06.004 · Cited by 25
Neurosurgical gene therapy for central nervous system diseases
Patel, R. V., Nanda, P., & Richardson, R. M. (2024) · Neurotherapeutics · doi:10.1016/j.neurot.2024.e00434 · Cited by 23
“Dual AAV vector approaches in which smaller transgenes can recombine to form a larger transgene are also being explored but have not made it to clinical trials for a neurological indication [86].”
BackgroundAAV Vector-Mediated Antibody Delivery (A-MAD) in the Central Nervous System
Marino, M., & Holt, M. G. (2022) · Frontiers in Neurology · doi:10.3389/fneur.2022.870799 · Cited by 24
“2 kb is inefficient, and unfavorably impacts on transduction (80).”
BackgroundRevolution of AAV in Drug Discovery: From Delivery System to Clinical Application
Yin, L., He, H., Zhang, H., Shang, Y., Fu, C., Wu, S., & Jin, T. (2025) · Journal of Medical Virology · doi:10.1002/jmv.70447 · Cited by 23
rAAV-Mediated Cochlear Gene Therapy: Prospects and Challenges for Clinical Application
Blanc, F., Mondain, M., Bemelmans, A., Affortit, C., Puel, J., & Wang, J. (2020) · Journal of Clinical Medicine · doi:10.3390/jcm9020589 · Cited by 21
Abulimiti, A., Lai, M. S., & Chang, R. C. (2021) · Mechanisms of Ageing and Development · doi:10.1016/j.mad.2021.111549 · Cited by 23
Maturana, C. J., Verpeut, J. L., Pisano, T. J., Dhanerawala, Z. M., Esteves, A. D., Enquist, L. W., & Engel, E. A. (2020) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2020.04.004 · Cited by 19
“9 kb.(43) For example, CNS therapies for Pompe disease(44) and Parkinson’s disease(45) are based on delivery of relatively large genes such as GAA (2.”
BackgroundPhelps, M., & Balazs, A. B. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.734304 · Cited by 21
“Following transduction, these recombinant viral genomes form head-to-tail concatemers within the nucleus, where they persist as non-integrated episomal genomes (105, 106).”
BackgroundCRISPR genome engineering for retinal diseases
Kantor, A., McClements, M. E., Peddle, C. F., Fry, L. E., Salman, A., Cehajic‐Kapetanovic, J., Xue, K., & MacLaren, R. E. (2021) · Progress in molecular biology and translational science · doi:10.1016/bs.pmbts.2021.01.024 · Cited by 19
Viral Vectors, Engineered Cells and the CRISPR Revolution
DiCarlo, J. E., Deeconda, A., & Tsang, S. H. (2017) · Advances in experimental medicine and biology · doi:10.1007/978-3-319-63904-8_1 · Cited by 19
Assessment of AAV Dual Vector Safety in the Abca4−/− Mouse Model of Stargardt Disease
McClements, M. E., Barnard, A. R., Charbel Issa, P., & MacLaren, R. E. (2020) · Translational Vision Science & Technology · doi:10.1167/tvst.9.7.20 · Cited by 18
“10,11 Briefly, the overlapping dual vector approach requires that the basic elements of an AAV gene therapy transgene be split across two separate transgenes.”
Allen, D. G., Kalter, N., Rosenberg, M., & Hendel, A. (2023) · Pharmaceutics · doi:10.3390/pharmaceutics15051329 · Cited by 18
Chamberlain, K., Riyad, J. M., Garnett, T., Kohlbrenner, E., Mookerjee, A., Elmastour, F., Bénard, L., Chen, J., VandenDriessche, T., Chuah, M., et al. (2018) · Human Gene Therapy · doi:10.1089/hum.2017.188 · Cited by 17
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Methods for CRISPR-Cas as Ribonucleoprotein Complex Delivery In Vivo
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“cases, as the CDS reconstitution is dependent on complex DNA repair pathways and can vary from different tissue/cell types, leading to diverse gene reconstitution rates [2,6].”
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“However, this is not a major concern for most hearing-related applications, with a median human protein size of 375 amino acids [63], since the cDNA of about 78 % of the proteins associated with hereditary hearing loss are under the size limit of the”
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“Since the large size of the SCN1A gene (cDNA ~6 kb) precludes packaging into AAV vectors, AAV-mediated gene replacement therapy is not feasible for SCN1A [92].”
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“As a result of this the superinfection, in the event of the presence of helper viruses, cannot appear as rAAVs are not able to integrate into the wtAAV’s preferential integration and trigger long-term transgene expression in postmitotic tissues, most likely because they persist as episomes within the host cell [59].”
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“The solution strategy that has been put forward to solve this problem is the use of new site-mutated AAVs.”
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“113 For example, nanoparticles may not be able to penetrate the layers of tissue in the eye to reach the target cells and have high toxicity.”
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“…partial capsids in AAV formulations is that they do not help to improve overall transduction because they contain no therapeutic GOI and could inhibit AAVs containing GOI (e.g., via competition for target receptors, co-receptor recognition, or binding saturation) to reach the target cells (45–53).”
Viral Vectors for Gene Therapy
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Synthetic genomics for curing genetic diseases
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Metadata
- Authors
- Chamberlain, K., Riyad, J. M., & Weber, T.
- Affiliations
- Icahn School of Medicine at Mount Sinai (US)
- Corresponding author
- Weber, Thomas
- Venue
- Human Gene Therapy Methods
- Publisher
- SAGE Publications
- Published
- 2016-02
- Volume
- 27
- Issue
- 1
- Pages
- 1-12
- ISSN
- 1946-6536 (ISSN-L)
- 1946-6544
- PMC
- PMC4761816
- OpenAlex
- W2280525353
- Semantic Scholar
- 2025d56e2cca1328dc2aed3bfe2dab7881704456
- Cited by
- 141
- References
- 74
- Citations by year
- 2026: 10 · 2025: 10 · 2024: 8 · 2023: 13 · 2022: 17 · 2021: 22 · 2020: 20 · 2019: 12 · 2018: 13 · 2017: 12 · 2016: 4
- FWCI
- 1.83
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Human Gene Therapy Methods (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Fondation Leducq (14CVD03)
- National Institutes of Health (HL112324)
- National Heart, Lung, and Blood Institute (P50HL 112324, HL117505, R01HL117505, HL112324)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- recombinant adeno-associated virus vectors, co-infection, transgene expression, gene therapy vectors, transduction efficiency
- MeSH terms
- Dependovirus, Animals, DNA, Viral, Genetic Vectors, Humans, Genetic Therapy, Gene Expression, Genome, Viral, Gene Transfer Techniques, Virus Assembly, Transgenes, Capsid Proteins, Genome Size
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