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A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice
Open accessAbstract
Many genetic liver diseases in newborns cause repeated, often lethal, metabolic crises. Gene therapy using nonintegrating viruses such as adeno-associated virus (AAV) is not optimal in this setting because the nonintegrating genome is lost as developing hepatocytes proliferate. We reasoned that newborn liver may be an ideal setting for AAV-mediated gene correction using CRISPR-Cas9. Here we intravenously infuse two AAVs, one expressing Cas9 and the other expressing a guide RNA and the donor DNA, into newborn mice with a partial deficiency in the urea cycle disorder enzyme, ornithine transcarbamylase (OTC). This resulted in reversion of the mutation in 10% (6.7-20.1%) of hepatocytes and increased survival in mice challenged with a high-protein diet, which exacerbates disease. Gene correction in adult OTC-deficient mice was lower and accompanied by larger deletions that ablated residual expression from the endogenous OTC gene, leading to diminished protein tolerance and lethal hyperammonemia on a chow diet.
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“As a result, in applications calling for corrective gene insertion, a second AAV vector encoding the sgRNA and/or a template sequence for homologydirected DNA repair must be used, reducing efficiency due to the need for cells to acquire both AAV vectors at once 59,60 .”
From The promise and challenge of therapeutic genome editing (2020) · cites this paper as [59]
Experimental — accuracy not yet measuredIn the citing paper
“Currently, researchers most frequently employ a dual AAV system for CRISPR-mediated gene correction: one AAV expresses Cas9 while the other expresses the sgRNA and donor template DNA [24].”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [24]
In the citing paper
“Furthermore, there was an increase in survival of mice with a high-protein diet, a stressor which exacerbates this in-born error of metabolism.”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [24]
In the citing paper
“After AAV delivery, these mice experienced a reversion of the mutation in 10% of their hepatocytes.”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [24]
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Citations
Cites
Genome engineering using the CRISPR-Cas9 system
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Items 201–300 of 564
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Self-citationLuo, W., Galvan, D. L., Woodard, L. E., Dorset, D. C., Levy, S. A., & Wilson, M. H. (2017) · Nucleic Acids Research · doi:10.1093/nar/gkx572 · Cited by 42
Personalised genome editing – The future for corneal dystrophies
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“This dual-AAV system has been demonstrated in mouse brain in vivo to study gene function of Mecp2 and in mouse liver in vivo where gene correction of Otc was demonstrated (Swiech et al., 2015; Yang et al., 2016).”
BackgroundMethodNanotechnology-enabled gene delivery for cancer and other genetic diseases
Jiang, T., Gonzalez, K. M., Cordova, L. E., & Lu, J. (2023) · Expert Opinion on Drug Delivery · doi:10.1080/17425247.2023.2200246 · Cited by 45
“[141] delivered SaCas9 and repaired DNA template”
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AAV capsid prioritization in normal and steatotic human livers maintained by machine perfusion
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“In addition, DNA recombination-dependent strategies, such as delivery of large genes with dual AAV vectors 79 and Cas9-mediated gene editing using templates delivered with AAV vectors 80 , will probably be less effective in these individuals.”
CRISPR/Cas9: at the cutting edge of hepatology
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Therapeutic genome editing in cardiovascular diseases
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Paving the way towards precise and safe CRISPR genome editing
Śledziński, P., Dabrowska, M., Nowaczyk, M., & Olejniczak, M. (2021) · Biotechnology Advances · doi:10.1016/j.biotechadv.2021.107737 · Cited by 43
State‐of‐the‐art 2023 on gene therapy for phenylketonuria
Martinez, M., Harding, C. O., Schwank, G., & Thöny, B. (2023) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12651 · Cited by 40
“This low repair frequency is similar to that achieved in mouse models of OTC deficiency, 88 hereditary tyrosinemia, 89 and hemophilia B. 90 However, if the PKU mice were treated with vanillin, a potent NHEJ inhibitor, 91 after rAAV vector delivery, the frequency of HDR increased to 13%.”
Tycko, J., Barrera, L. A., Huston, N. C., Friedland, A. E., Wu, X., Gootenberg, J. S., Abudayyeh, O. O., Myer, V. E., Wilson, C. J., & Hsu, P. D. (2018) · Nature Communications · doi:10.1038/s41467-018-05391-2 · Cited by 38
Curative Ex Vivo Hepatocyte-Directed Gene Editing in a Mouse Model of Hereditary Tyrosinemia Type 1
VanLith, C. J., Guthman, R. M., Nicolas, C. T., Allen, K. L., Du, Z., Joo, D. J., Nyberg, S. L., Lillegard, J. B., & Hickey, R. (2018) · Human Gene Therapy · doi:10.1089/hum.2017.252 · Cited by 39
“This obstacle has been previously highlighted with recent in vivo gene-editing approaches, with DNA repair efficiencies significantly lower in adult mice compared to neonatal mice.(43) A partial hepatectomy was utilized as a clinically relevant method to improve gene editing in transplanted hepatocytes.”
MethodCRISPR/Cas9 Delivery Mediated with Hydroxyl‐Rich Nanosystems for Gene Editing in Aorta
Zhang, X., Chen, X., Gao, S., Li, P., Kong, Y., Li, T., Li, Y., Xu, F., & Du, J. (2019) · Advanced Science · doi:10.1002/advs.201900386 · Cited by 40
Immunity to Cas9 as an Obstacle to Persistent Genome Editing
Gough, V., & Gersbach, C. A. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.05.007 · Cited by 41
Cheng, Y., Wang, H., & Li, M. (2023) · Journal of Diabetes and its Complications · doi:10.1016/j.jdiacomp.2023.108524 · Cited by 45
CRISPR/Cas9 as a therapeutic tool for triple negative breast cancer: from bench to clinics
Tiwari, P. K., Ko, T.-H., Dubey, R., Chouhan, M., Tsai, L.-W., Singh, H. N., Chaubey, K. K., Dayal, D., Chiang, C., & Kumar, S. (2023) · Frontiers in Molecular Biosciences · doi:10.3389/fmolb.2023.1214489 · Cited by 42
“CRISPR/Cas9 may also produce genoand cellular toxicity if delivered via the two current methods such as viral and bacteriophage-derived vectors (Yang et al., 2016).”
BackgroundYi, Z., Zhao, Y., Yi, Z., Zhang, Y., Tang, G.-B., Zhang, X., Tang, H., Zhang, W., Zhao, Y., Xu, H., et al. (2023) · Genome biology · doi:10.1186/s13059-023-03086-6 · Cited by 41
“Although direct repair of mutations causing diseases is possible through gene-editing enzymes [4, 5], the ectopic expression of these enzymes is often associated with non-negligible problems, including extensive global off-target of genomic DNA and/or RNA transcripts [6–9], immunogenicity [10–12],…”
BackgroundMusunuru, K. (2017) · Journal of the American College of Cardiology · doi:10.1016/j.jacc.2017.10.002 · Cited by 38
Self-citationAAV-Mediated Gene Delivery to the Liver: Overview of Current Technologies and Methods
Palaschak, B., Herzog, R. W., & Markusic, D. M. (2019) · Methods in molecular biology · doi:10.1007/978-1-4939-9139-6_20 · Cited by 37
Progress and challenges in development of new therapies for urea cycle disorders
Soria, L. R., Mew, N. A., & Brunetti‐Pierri, N. (2019) · Human Molecular Genetics · doi:10.1093/hmg/ddz140 · Cited by 38
A Robust System for Production of Superabundant VP1 Recombinant AAV Vectors
Qizhao, W., Wu, Z., Zhang, J., Firrman, J., Wei, H., Zhuang, Z., Liu, L., Miao, L., Hu, Y., Li, D., et al. (2017) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2017.11.002 · Cited by 37
Tso, F. Y., West, J. T., & Wood, C. A. (2019) · Journal of Virology · doi:10.1128/jvi.02183-18 · Cited by 38
mRNA and gene editing: Late breaking therapies in liver diseases
Zabaleta, N., Torella, L., Weber, N. D., & González‐Aseguinolaza, G. (2022) · Hepatology · doi:10.1002/hep.32441 · Cited by 39
“[66,67] Proof- of- concept studies have also been carried out for OCTD,[68] hemophilia B,[69] and phenylketonuria (PKU).”
BackgroundIn vivo therapeutic genome editing via CRISPR/Cas9 magnetoplexes for myocardial infarction
Park, H., Kim, D., Cho, B., Byun, J., Kim, Y. S., Ahn, Y., Hur, J., Oh, Y., & Kim, J. (2021) · Biomaterials · doi:10.1016/j.biomaterials.2021.121327 · Cited by 37
Advances in CRISPR Delivery Methods: Perspectives and Challenges
Demirci, S., Essawi, K., Germino-Watnick, P., Liu, X., Hakami, W., & Tisdale, J. F. (2022) · The CRISPR Journal · doi:10.1089/crispr.2022.0051 · Cited by 37
“77,82 However, as AAVs have a limited payload ( 4.7 kb), it makes it challenging to package the commonly used Streptococcus pyogenes (Sp)Cas9 and a gRNA together ( * 4.2 kb).”
InfluentialGene therapy for glycogen storage diseases
Koeberl, D. D., Koch, R. L., Lim, J., Brooks, E. D., Arnson, B., Sun, B., & Kishnani, P. S. (2023) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12654 · Cited by 39
CRISPR/Cas9-Mediated Gene Correction in Newborn Rabbits with Hereditary Tyrosinemia Type I
Li, N., Gou, S., Wang, J., Zhang, Q., Huang, X., Xie, J., Li, L., Jin, Q., Ouyang, Z., Chen, F., et al. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.11.023 · Cited by 38
Beclin‐1‐mediated activation of autophagy improves proximal and distal urea cycle disorders
Soria, L. R., Gurung, S., Sabbata, G. D., Perocheau, D. P. B., Angelis, A. D., Bruno, G., Polishchuk, E., Paris, D., Cuomo, P., Motta, A., et al. (2020) · EMBO Molecular Medicine · doi:10.15252/emmm.202013158 · Cited by 34
CRISPR/Cas-Dependent and Nuclease-Free In Vivo Therapeutic Gene Editing
Dasgupta, I., Flotte, T. R., & Keeler, A. M. (2021) · Human Gene Therapy · doi:10.1089/hum.2021.013 · Cited by 38
“Some of the in vivo gene therapy studies revealed minimal or no off-target editing at the predicted sites, which is reassuring.(58,69,70,145) However, the possibility of off-target editing beyond the predicted sites requires the design of an unbiased genome-wide sequencing method.”
BackgroundCorneal Regeneration Using Gene Therapy Approaches
Sarkar, S., Panikker, P., D’Souza, S., Shetty, R., Mohan, R. R., & Ghosh, A. (2023) · Cells · doi:10.3390/cells12091280 · Cited by 36
“To address this issue, a dualAAV vector design has been implemented in which the Cas9 nuclease and the single-guide RNA (sgRNA) cassettes have been packaged in two separate vectors and delivered to the target tissue, which showed highly efficient genome editing in the hepatocyte cells [168].”
MethodUnlocking Genome Editing: Advances and Obstacles in CRISPR/Cas Delivery Technologies
Kaupbayeva, B., Tsoy, A., Safarova, Y., Nurmagambetova, A., Murata, H., Matyjaszewski, K. A., & Askarova, S. (2024) · Journal of Functional Biomaterials · doi:10.3390/jfb15110324 · Cited by 40
“Yang et al. illustrated the use of a dual AAV system to deliver the CRISPR/Cas9 technology, utilizing one vector to introduce SaCas9 and another to provide sgRNA targeting ornithine transcarbamylase (OTC) in mice with partial OTC deficiency, which affects the urea cycle in the liver [19].”
Split Cas9, Not Hairs − Advancing the Therapeutic Index of CRISPR Technology
Schmelas, C., & Grimm, D. (2018) · Biotechnology Journal · doi:10.1002/biot.201700432 · Cited by 35
Chen, D., Xu, T., Tu, M., Xu, J., Zhou, C., Cheng, L., Yang, R., Yang, T., Zheng, W., He, X., et al. (2018) · Frontiers in Molecular Neuroscience · doi:10.3389/fnmol.2017.00453 · Cited by 35
“Genome editing based gene therapy would be a great promising method for treating genetic diseases (Yin et al., 2014; Suzuki et al., 2016; Yang et al., 2016).”
MethodGenetic-Based Approaches to Inherited Metabolic Liver Diseases
Zabaleta, N., Hommel, M., Salas, D., & González‐Aseguinolaza, G. (2019) · Human Gene Therapy · doi:10.1089/hum.2019.140 · Cited by 35
“170 The percentage of corrected hepatocytes in the absence of a growth advantage was higher in mice treated as neonates than in mice treated as adults and resulted in a greater therapeutic effect.”
Khatib, T. Z., Osborne, A., Yang, S., Ali, Z., Jia, W., Manyakin, I., Hall, K., Watt, R., Widdowson, P. S., & Martin, K. R. G. (2021) · Science Advances · doi:10.1126/sciadv.abd2590 · Cited by 35
“However, dual promoter techniques have previously shown selective expression ( 16 ) and reduced efficiency ( 17 ).”
Targeting mechanosensitive endothelial TXNDC5 to stabilize eNOS and reduce atherosclerosis in vivo
Yeh, C.-F., Cheng, S.-H., Lin, Y.-S., Shentu, T.-P., Huang, R.-T., Zhu, J., Chen, Y.-T., Kumar, S., Lin, M.-S., Kao, H.-L., et al. (2022) · Science Advances · doi:10.1126/sciadv.abl8096 · Cited by 44
“…AAV system with tissue-specific promoter or tropism modification was used to overcome these barriers; however, the in vivo efficiency of dual AAV system was lower compared to single AAV approaches because of the heterogeneity of cotransduction, especially when administered systemically ( 57 , 58 ).”
Chen, Y., Chen, X., Wu, D., Xin, H., Chen, D., Li, D., Pan, H., Zhou, C., & Yuan, P. (2020) · Chemistry of Materials · doi:10.1021/acs.chemmater.0c02571 · Cited by 35
Haley, R. M., Padilla, M. S., El-Mayta, R. D., Joseph, R. A., Weber, J. A., Figueroa‐Espada, C. G., Mukalel, A. J., Ricciardi, A. S., Palanki, R., Geisler, H. C., et al. (2025) · ACS Nano · doi:10.1021/acsnano.4c16617 · Cited by 38
CRISPR applications in ophthalmologic genome surgery
Cabral, T., DiCarlo, J. E., Justus, S., Sengillo, J. D., Xu, Y., & Tsang, S. H. (2017) · Current Opinion in Ophthalmology · doi:10.1097/icu.0000000000000359 · Cited by 33
TRAP-seq identifies cystine/glutamate antiporter as a driver of recovery from liver injury
Wang, A. W., Wangensteen, K. J., Wang, Y. J., Zahm, A. M., Moss, N., Noam Erez, A., & Kaestner, K. H. (2018) · Journal of Clinical Investigation · doi:10.1172/jci95120 · Cited by 30
“We coexpressed FAH with either 10 single-guide RNAs (sgRNAs) targeting Slc7a11 exons (Fah-sgSlc7a11) or 10 control sgRNAs targeting luciferase (Fah-gCtl) and performed hydrodynamic tail-vein injection of these sgRNAs, together with adeno-associated virus 8 (AAV8) expressing Staphylococcus aureus Cas9 (SaCas9) to allow…”
MethodVanLith, C. J., Guthman, R. M., Nicolas, C. T., Allen, K. L., Liu, Y., Chilton, J. A., Tritz, Z. P., Nyberg, S. L., Kaiser, R. A., Lillegard, J. B., et al. (2019) · Hepatology Communications · doi:10.1002/hep4.1315 · Cited by 32
Using CRISPR/Cas9 to model human liver disease
Alves‐Bezerra, M., Furey, N., Johnson, C. G., & Bissig, K. (2019) · JHEP Reports · doi:10.1016/j.jhepr.2019.09.002 · Cited by 32
CRISPR-Cas9 Genome Editing for Treatment of Atherogenic Dyslipidemia
Chadwick, A. C., & Musunuru, K. (2017) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.117.309326 · Cited by 33
“To date, the most successful execution of HDR in a mouse model (ornithine transcarbamylase deficiency) resulted in 10% to 20% allele correction; notably this level of correction was observed in newborn mice with rapidly proliferating hepatocytes but not in adult mice with relatively static hepatocytes in which only 1%…”
Self-citationDesign of efficacious somatic cell genome editing strategies for recessive and polygenic diseases
Carlson-Stevermer, J., Das, A., Abdeen, A. A., Fiflis, D., Grindel, B. I., Saxena, S., Akcan, T., Alam, T., Kletzien, H., Kohlenberg, L., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-20065-8 · Cited by 32
Shakirova, A. I., Karpov, T. E., Komarova, Y., & Lepik, K. V. (2023) · Frontiers in Genome Editing · doi:10.3389/fgeed.2023.1068637 · Cited by 33
“Such properties of AAVs have formed the basis for their domination as donor vectors both in vitro and in vivo (Table 2) (Sather et al., 2015; Yang et al., 2016; Yin et al., 2016; Pattabhi et al., 2019; Lattanzi et al., 2021; Wilkinson et al., 2021).”
BackgroundThe technical risks of human gene editing
Davies, B. (2019) · Human Reproduction · doi:10.1093/humrep/dez162 · Cited by 37
“%) of disruptive large deletions was found (Yang et al., 2016).”
BackgroundEfficient in vivo editing of OTC-deficient patient-derived primary human hepatocytes
Ginn, S. L., Amaya, A. K., Liao, S. H. Y., Zhu, E., Cunningham, S. C., Lee, M., Hallwirth, C. V., Logan, G. J., Tay, S. S., Cesare, A. J., et al. (2019) · JHEP Reports · doi:10.1016/j.jhepr.2019.100065 · Cited by 32
“Only a single study reports the use of dual AAV-based editing reagents in vivo to treat ornithine transcarbamylase (OTC) deficiency, in a murine model with mild disease.(18) Although this work provides important proof-of-concept data, sequence differences between mouse and man prevent the clinical translation of these…”
BackgroundYin, S., Ma, L., Shao, T., Zhang, M., Guan, Y., Wang, L., Hu, Y., Chen, X., Han, H., Shen, N., et al. (2020) · Science China Life Sciences · doi:10.1007/s11427-020-1744-6 · Cited by 33
Self-citationPre-clinical non-viral vectors exploited for in vivo CRISPR/Cas9 gene editing: an overview
Rouatbi, N., McGlynn, T., & Al‐Jamal, K. T. (2022) · Biomaterials Science · doi:10.1039/d1bm01452h · Cited by 33
Regenerative medicine: postnatal approaches
Tam, P. K. H., Wong, K. K. Y., Atala, A., Giobbe, G. G., Booth, C., Gruber, P. J., Mononen, M., Rafii, S., Rando, T. A., Vacanti, J. P., et al. (2022) · The Lancet Child & Adolescent Health · doi:10.1016/s2352-4642(22)00193-6 · Cited by 33
Engineered CRISPR Systems for Next Generation Gene Therapies
Pineda, M., Moghadam, F., Ebrahimkhani, M. R., & Kiani, S. (2017) · ACS Synthetic Biology · doi:10.1021/acssynbio.7b00011 · Cited by 30
“33 (2), 187−97. (58) Tsai, S. Q., Nguyen, N. T., Malagon-Lopez, J., Topkar, V. V., Aryee, M. J., and Joung, J. K. (2017) CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR-Cas9 nuclease off-targets. Nat. Methods 14, 607. (59) Davis, A. J., and Chen, D. J. (2013) DNA double strand break repair via no…”
InfluentialWang, X., Raghavan, A., Peters, D., Pashos, E. E., Rader, D. J., & Musunuru, K. (2017) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.117.310103 · Cited by 29
Self-citationSabatini, P. V., Wang, J., Rupp, A. C., Affinati, A. H., Flak, J. N., Li, C., Olson, D. P., & Myers, M. G. (2021) · eLife · doi:10.7554/elife.66835 · Cited by 29
“Based upon our results, and since dual AAVs have been used successfully previously (Akil et al., 2019; Xu et al., 2018; Yang et al., 2016) and transduce the same cells in vivo (Vardy et al.”
Elsharkasy, O. M., Hegeman, C. V., Driedonks, T. A. P., Liang, X., Lansweers, I., Cotugno, O. L., de Groot, I. Y., de Wit, Z. E. M. N. J., Garcia-Guerra, A., Moorman, N. J. A., et al. (2025) · Nature Communications · doi:10.1038/s41467-025-65995-3 · Cited by 37
“To this end, various intracellular delivery strategies have been employed to facilitate Cas9 RNP delivery, including viral vectors, lipid nanoparticles (LNPs), polymers, and cell penetrating peptides (CPPs) 10 – 13 .”
Gene therapy for inherited metabolic diseases
Yılmaz, B. Ş., Gurung, S., Perocheau, D. P. B., Counsell, J. R., & Baruteau, J. (2020) · PubMed Central · doi:10.34763/jmotherandchild.20202402si.2004.000009 · Cited by 31
Gene therapy for urea cycle defects: An update from historical perspectives to future prospects
Duff, C. E., Alexander, I. E., & Baruteau, J. (2023) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12609 · Cited by 30
“88 Another approach combining DNA double-strand break and homologous recombination via two 900 bp-long homologous arms flanking the OTC donor template was then successfully tested using the same sgRNA via a similar dual AAV approach injected systemically in neo-natal OTCD Spf ash pups.”
MethodArnaoutova, I., Zhang, L., Chen, H., Mansfield, B. C., & CHOU, J. Y. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.12.027 · Cited by 30
“Using the G6pc-R83C mice, we explore the efficacy of CRISPR/Cas9 to correct metabolic abnormalities in these mice using a dual AAV strategy that targets primarily the liver.(11) We used two different virally delivered doses of the editing reagents: AAV-H and AAV-L.”
MethodStrategies to improve safety profile of AAV vectors
Suoranta, T., Laham-Karam, N., & Ylä‐Herttuala, S. (2022) · Frontiers in Molecular Medicine · doi:10.3389/fmmed.2022.1054069 · Cited by 32
“Combinational strategies with CRISPR-Cas9 have also been used for example in vitro stem cell editing and also in vivo (Dever et al., 2016; Yang et al., 2016).”
MethodCRISPR/Cas9 system: a powerful technology for in vivo and ex vivo gene therapy
Zhang, X., Wang, L., Liu, M., & Li, D. (2017) · Science China Life Sciences · doi:10.1007/s11427-017-9057-2 · Cited by 28
“The saCas9 and sgRNA-donor were delivered by two AAV vectors, through temporal vein injection (Yang et al., 2016).”
MethodSelf-citationGinocchio, V. M., Ferla, R., Auricchio, A., & Brunetti‐Pierri, N. (2019) · Human Gene Therapy · doi:10.1089/hum.2019.151 · Cited by 29
“52 The efficiency of NHEJ has been exploited to insert a donor DNA with the therapeutic transgene at a given locus in the absence of homology arms.”
Jiang, X., Shao, M., Liu, X., Liu, X., Zhang, X., Wang, Y., Yin, K., Wang, S., Hu, Y., José, P. A., et al. (2021) · Advanced Science · doi:10.1002/advs.202003706 · Cited by 27
Versatile enzymology and heterogeneous phenotypes in cobalamin complementation type C disease
Esser, A. J., Mukherjee, S., Dereven’kov, I. А., Makarov, S. V., Jacobsen, D. W., Spiekerkoetter, U., & Hannibal, L. (2022) · iScience · doi:10.1016/j.isci.2022.104981 · Cited by 28
“…2015), hereditary tyrosinemia type I (Nygaard et al., 2016; Pankowicz et al., 2016; Yin et al., 2016, 2014), ornithine transcarbamylase deficiency (Yang et al., 2016), lysosomal storage disorders (Sharma et al., 2015), and glycogen storage disease (Landau et al., 2016) showing successful…”
Homology-directed repair of an MYBPC3 gene mutation in a rat model of hypertrophic cardiomyopathy
Nie, J., Han, Y., Jin, Z., Hang, W., Shu, H., Zheng, W., Ni, L., & Wang, D. W. (2023) · Gene Therapy · doi:10.1038/s41434-023-00384-3 · Cited by 28
“Prior studies have shown that HDR has great potential for the treatment of genetic diseases such as hereditary tyrosinemia [17], OTC deficiency [9], and inherited blindness [18].”
BackgroundAdvances in understanding disease mechanisms and potential treatments for Crigler–Najjar syndrome
Bortolussi, G., & Muro, A. F. (2018) · Expert Opinion on Orphan Drugs · doi:10.1080/21678707.2018.1495558 · Cited by 27
Correction of a urea cycle defect after ex vivo gene editing of human hepatocytes
Zabulica, M., Srinivasan, R. C., Akçakaya, P., Allegri, G., Bestas, B., Firth, M. A., Hammarstedt, C., Jakobsson, T., Jakobsson, T., Ellis, E. C. S., et al. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.01.024 · Cited by 29
“The elegant studies of Yang et al.12 described the use of in vivo dual-adeno-associated virus (AAV)-delivered CRISPR gene editing components to repair the OTC gene in the sparse fur ash (spfash) rodent model of OTC deficiency.”
BackgroundKumari, R., Saha, T., Kumar, P., & Singh, A. K. (2024) · Physiology and Molecular Biology of Plants · doi:10.1007/s12298-024-01486-x · Cited by 27
The Yin and Yang of nucleic acid-based therapy in the brain
Gustincich, S., Zucchelli, S., & Mallamaci, A. (2016) · Progress in Neurobiology · doi:10.1016/j.pneurobio.2016.11.001 · Cited by 27
CRISPR/Cas based gene editing: marking a new era in medical science
Prasad, K. M., George, A., Ravi, N. S., & Mohankumar, K. M. (2021) · Molecular Biology Reports · doi:10.1007/s11033-021-06479-7 · Cited by 27
Laemmle, A., Poms, M., Hsu, B. Y., Borsuk, M., Rüfenacht, V., Robinson, J. F., Sadowski, M. C., Nuoffer, J., Häberle, J., & Willenbring, H. (2021) · Hepatology · doi:10.1002/hep.32247 · Cited by 27
“One such approach could be gene addition or gene editing, and indeed adeno-associated viruses were shown to correct OTCD in preclinical animal models [38–40] and are currently being investigated in clinical trials in adult human subjects suffering from mild OTCD (NCT02991144).”
Gu, X. G., Hu, X., Wang, D., Xu, Z., Wang, F., Li, D., Li, G., Yang, H., Li, H., Zuo, E., et al. (2022) · Cell Research · doi:10.1038/s41422-022-00624-y · 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
Therapeutic genome engineering via CRISPR ‐Cas systems
Moreno, A. M., & Mali, P. (2017) · WIREs Systems Biology and Medicine · doi:10.1002/wsbm.1380 · Cited by 25
“The variable spacer region is typically a short ~20 bp sequence that is complementary to the target loci, and must be flanked at the 30 end by a conserved protospacer adjacent motif (PAM) sequence, for instance, NGG in SpCas9.”
ResultInnovative Precision Gene‐Editing Tools in Personalized Cancer Medicine
Dai, X., Blancafort, P., Wang, P., Sgro, A., Thompson, E. W., & Ostrikov, K. (2020) · Advanced Science · doi:10.1002/advs.201902552 · Cited by 28
Optical Manipulation of CRISPR/Cas9 Functions: From Ultraviolet to Near-Infrared Light
Chen, Y., Yan, X., & Yuan, P. (2020) · ACS Materials Letters · doi:10.1021/acsmaterialslett.0c00041 · Cited by 26
The landscape of CRISPR/Cas9 for inborn errors of metabolism
Leal, A. F., Nidhi, F., Benincore-Flórez, E., Herreño-Pachón, A. M., Echeverri-Peña, O. Y., Alméciga-Díaz, C. J., & Tomatsu, S. (2022) · Molecular Genetics and Metabolism · doi:10.1016/j.ymgme.2022.106968 · Cited by 27
Genetic Therapy Approaches for Ornithine Transcarbamylase Deficiency
Yılmaz, B. Ş., & Gissen, P. (2023) · Biomedicines · doi:10.3390/biomedicines11082227 · Cited by 26
“However, gene correction was significantly lower in the adult mice with a reduced protein tolerance and fatal hyperammonemia on a chow diet [89].”
BackgroundHu, J., Liu, N., Song, D. H., Steer, C. J., Zheng, G., & Song, G. (2023) · Oncogene · doi:10.1038/s41388-023-02757-9 · Cited by 26
“In sum, by disrupting the positive feedback loop between the cholesterol synthesis and the PPP, miR-206 inhibited growth of HCC.”
BackgroundCRISPR Editing in Biological and Biomedical Investigation
Ju, X., Xu, J., & Sun, Z. S. (2017) · Journal of Cellular Biochemistry · doi:10.1002/jcb.26154 · Cited by 27
“Various other preclinical studies have shown that highly specific CRISPR therapeutics can be implemented for other liver diseases, such as reversion of the ornithine transcarbamylase (OTC) gene mutation in hyperammonemia [Yang et al., 2016].”
BackgroundTherapeutic applications of CRISPR RNA-guided genome editing
Koo, T., & Kim, J. (2016) · Briefings in Functional Genomics · doi:10.1093/bfgp/elw032 · Cited by 24
Jiang, W., Liu, L., Chang, Q., Xing, F., Ma, Z., Fang, Z., Zhou, J., Fu, L., Huiyang, W., Huang, X., et al. (2018) · Scientific Reports · doi:10.1038/s41598-018-19774-4 · Cited by 24
Multiplexed promoterless gene expression with CRISPReader
Zhan, H., Zhou, Q. Y., Gao, Q., Li, J., Huang, W., & Liu, Y. (2019) · Genome biology · doi:10.1186/s13059-019-1712-5 · Cited by 25
Recent advances in therapeutic CRISPR-Cas9 genome editing: mechanisms and applications
Zhou, L., & Yao, S. (2023) · Molecular Biomedicine · doi:10.1186/s43556-023-00115-5 · Cited by 29
Innovations in CRISPR technology
Brooks, A. K., & Gaj, T. (2018) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2018.03.007 · Cited by 26
Versatile High-Throughput Fluorescence Assay for Monitoring Cas9 Activity
Seamon, K. J., Light, Y. K., Saada, E. A., Schoeniger, J. S., & Harmon, B. (2018) · Analytical Chemistry · doi:10.1021/acs.analchem.8b01155 · Cited by 24
Allegri, G., Deplazes, S., Rimann, N., Causton, B., Scherer, T., Leff, J. W., Díez‐Fernández, C., Klimovskaia, A., Fingerhut, R., Krijt, J., et al. (2019) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12068 · Cited by 24
Lotfinia, M., Abdollahpour‐Alitappeh, M., Hatami, B., Zali, M. R., & Karimipoor, M. (2019) · Clinical and Experimental Medicine · doi:10.1007/s10238-019-00557-8 · Cited by 24
Utility of Self-Destructing CRISPR/Cas Constructs for Targeted Gene Editing in the Retina
Fan, L., Hung, S. S. C., Khalid, M. K. N. M., Wang, J.-H., Chrysostomou, V., Wong, V. H. Y., Singh, V., Wing, K., Tu, L., Bender, J. A., et al. (2019) · Human Gene Therapy · doi:10.1089/hum.2019.021 · Cited by 24
“29 Whether such mechanisms account for the functional deficits observed in our study requires further investigation.”
Gene editing in dermatology: Harnessing CRISPR for the treatment of cutaneous disease
Baker, C. M., & Hayden, M. S. (2020) · F1000Research · doi:10.12688/f1000research.23185.1 · Cited by 26
Induced Liver Regeneration Enhances CRISPR/Cas9-Mediated Gene Repair in Tyrosinemia Type 1
Zhang, Q., Tiyaboonchai, A., Nygaard, S. C., Baradar, K., Major, A. M., Balaji, N., & Grompe, M. (2020) · Human Gene Therapy · doi:10.1089/hum.2020.042 · Cited by 24
“Others employed a dual AAV system (one vector expressing Staphylococcus aureus Cas9 (saCas9) and the other containing homology donor) in a mouse model of ornithine transcarbamylase (OTC) deficiency.(9) Gene repair frequencies of 7–20% were seen in neonates, whereas much lower (0.”
BackgroundMethodResultInfluentialZhou, L., Su, J., Long, J., Tao, R. X., Tang, W., Qin, F., Liu, N., Wang, Y., Jiao, Y., Hu, Y. F., et al. (2022) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2022.01.001 · Cited by 24
“DISCUSSION Since the discovery of AAVs, much progress has been made in recent decades in the treatment of genetic diseases, especially liver-related metabolic disorders.(16,23,24) Compared with other viral vectors, rAAVs are relatively safe because they are less immunogenic and rarely integrate into the host genome.”
BackgroundSelf-citationBasic Principles and Clinical Applications of CRISPR-Based Genome Editing
Lim, J. M., & Kim, S. (2022) · Yonsei Medical Journal · doi:10.3349/ymj.2022.63.2.105 · Cited by 26
Chen, X., Niu, X., Liu, Y., Zheng, R., Yang, L., Lu, J., Yin, S., Wei, Y., Pan, J., Sayed, A. M. M., et al. (2022) · Journal of genetics and genomics/Journal of Genetics and Genomics · doi:10.1016/j.jgg.2022.06.001 · Cited by 25
Self-citationGene modification therapies for hereditary diseases in the fetus
Mattar, C. N. Z., Chan, J. K. Y., & Choolani, M. A. (2023) · Prenatal Diagnosis · doi:10.1002/pd.6347 · Cited by 23
Diagnostic and Management Issues in Patients with Late-Onset Ornithine Transcarbamylase Deficiency
Ibrahim, M., Gold, J. I., Woodall, A., Yılmaz, B. Ş., Gissen, P., & Stępień, K. M. (2023) · Children · doi:10.3390/children10081368 · Cited by 24
Oncolytic viral therapy for gliomas: Advances in the mechanisms and approaches to delivery
Romanishin, A. O., Vasilev, A., Khasanshin, E., Evtekhov, A., Pusynin, E., Rubina, K., Kakotkin, V., Agapov, M., & Semina, E. (2024) · Virology · doi:10.1016/j.virol.2024.110033 · Cited by 24
Genome Editing for the Study of Cardiovascular Diseases
Chadwick, A. C., & Musunuru, K. (2017) · Current Cardiology Reports · doi:10.1007/s11886-017-0830-5 · Cited by 28
Self-citationIn vivo gene therapy potentials of CRISPR-Cas9
Xue, H.-Y., Zhang, X., Wang, Y., Lu, X., Dai, W.-J., & Xu, Y. C. (2016) · Gene Therapy · doi:10.1038/gt.2016.25 · Cited by 24
Genome Editing in Neurosciences
Jaenisch, R., Gage, F. H., & Zhang, F. (2017) · Research and perspectives in neurosciences · doi:10.1007/978-3-319-60192-2 · Cited by 23
Metadata
- Authors
- Yang, Y., Wang, L., Bell, P., McMenamin, D., He, Z., White, J., Yu, H., Xu, C., Morizono, H., Musunuru, K., Batshaw, M. L., & Wilson, J. M.
- Affiliations
- Sichuan University (CN)
- University of Pennsylvania (US)
- Children's National (US)
- Brigham and Women's Hospital (US)
- Corresponding author
- Wilson, James M
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2016-02-01
- Volume
- 34
- Issue
- 3
- Pages
- 334-338
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC4786489
- OpenAlex
- W2253584908
- Cited by
- 556
- References
- 36
- Citations by year
- 2026: 20 · 2025: 25 · 2024: 36 · 2023: 52 · 2022: 55 · 2021: 61 · 2020: 70 · 2019: 62 · 2018: 72 · 2017: 71 · 2016: 30
- FWCI
- 63.21
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- Funders
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (P01HD057247)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- adeno-associated virus, CRISPR/Cas9, gene correction, ornithine transcarbamylase deficiency, urea cycle disorders, neonatal mice, liver disease, hyperammonemia, high-protein diet, hepatocytes, gene therapy
- MeSH terms
- Dependovirus, Animals, Disease Models, Animal, Genetic Vectors, Humans, Ornithine Carbamoyltransferase, Viruses, RNA Editing, Gene Transfer Techniques, Ornithine Carbamoyltransferase Deficiency Disease, Mice, CRISPR-Cas Systems
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