A Multifunctional AAV-CRISPR-Cas9 and Its Host Response
Open accessAbstract
CRISPR-Cas9 delivery by adeno-associated virus (AAV) holds promise for gene therapy but faces critical barriers on account of its potential immunogenicity and limited payload capacity. Here, we demonstrate genome engineering in postnatal mice using AAV-split-Cas9, a multifunctional platform customizable for genome editing, transcriptional regulation, and other previously impracticable applications of AAV-CRISPR-Cas9. We identify crucial parameters that impact efficacy and clinical translation of our platform, including viral biodistribution, editing efficiencies in various organs, antigenicity, immunological reactions, and physiological outcomes. These results reveal that AAV-CRISPR-Cas9 evokes host responses with distinct cellular and molecular signatures, but unlike alternative delivery methods, does not induce extensive cellular damage in vivo. Our study provides a foundation for developing effective genome therapeutics.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
An alternate method for expressing large proteins relying on intein protein sequences has been reported.
“An alternate method for expressing large proteins relying on intein protein sequences has also been reported (Li et al., 2008; Truong et al., 2015; Chew et al., 2016).”
From Evaluating Efficiencies of Dual AAV Approaches for Retinal Targeting (2017) · cites this paper as Chew et al., 2016
AAV is being developed as a delivery platform for in vivo gene editing in humans, raising the possibility of T cell responses against the editors, which are typically nucleases of bacterial origin.
“AAV is also being developed as a delivery platform for in vivo gene editing in humans, raising the possibility of T cell responses against the editors, which are typically nucleases of bacterial origin.20,21”
From TLR9-independent CD8+ T cell responses in hepatic AAV gene transfer through IL-1R1-MyD88 signaling (2024) · cites this paper as [20]
In Ai928 and WT mice, split-spCas9 packaged into two AAVs delivered by systemic and intramuscular injection induced production of specific antibodies to both AAV9 and Cas9, while only Cas9 induced a cellular immune response.
“Another study aimed at evaluating transduction efficiency and host response, delivered split-spCas9 packaged into two AAVs by systemic and intramuscular injection in Ai928 and WT (C57BL/6) mice.Serums were collected to measure anti-SpCas9 antibody levels, and muscle tissue was harvested to test the immune cells' profile by FACS.Interestingly, both AAV9 and Cas9 induced the production of specific”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [40]
Providing FK506 daily at 5 mg/kg reduced the number of activated CD4+ and CD8+ T-cells specific to Cas9 in mice.
“In a study aimed at evaluating the AAV-Cas9 immune responses in mice, providing FK506 daily at 5 mg/kg reduced the number of activated CD4 + and CD8 + T-cells that were specific to Cas9 [40].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [40]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2
Love, M. I., Huber, W., & Anders, S. (2014) · Genome Biology · doi:10.1186/s13059-014-0550-8 · Cited by 95,106 · see citing works
Cytoscape: A software Environment for integrated models of biomolecular interaction networks
Shannon, P., Markiel, A., Ozier, O., Baliga, N. S., Wang, J. T., Ramage, D., Amin, N., Schwikowski, B., & Ideker, T. (2003) · Genome Research · doi:10.1101/gr.1239303 · Cited by 51,888 · see citing works
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116 · see citing works
Trapnell, C., Williams, B. A., Pertea, G., Mortazavi, A., Kwan, G., van Baren, M. J., Salzberg, S. L., Wold, B. J., & Pachter, L. (2010) · Nature Biotechnology · doi:10.1038/nbt.1621 · Cited by 15,556 · see citing works
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339 · see citing works
RNA-Guided Human Genome Engineering via Cas9
Mali, P., Yang, L., Esvelt, K. M., Aach, J., Guell, M., DiCarlo, J. E., Norville, J. E., & Church, G. M. (2013) · Science · doi:10.1126/science.1232033 · Cited by 9,210 · see citing works
Self-citationA robust and high-throughput Cre reporting and characterization system for the whole mouse brain
Madisen, L., Zwingman, T. A., Sunkin, S. M., Oh, S. W., Zariwala, H. A., Gu, H., Ng, L. L., Palmiter, R. D., Hawrylycz, M. J., Jones, A. R., et al. (2010) · Nature Neuroscience · doi:10.1038/nn.2467 · Cited by 6,907 · see citing works
Bindea, G., Mlecnik, B., Hackl, H., Charoentong, P., Tosolini, M., Kirilovsky, A., Fridman, W.-H., Pagès, F., Trajanoski, Z., & Galon, J. (2009) · Bioinformatics · doi:10.1093/bioinformatics/btp101 · Cited by 6,280 · see citing works
CellProfiler: image analysis software for identifying and quantifying cell phenotypes
Carpenter, A. E., Jones, T. R., Lamprecht, M. R., Clarke, C., Kang, I. H., Friman, O., Guertin, D. A., Chang, J. H., Lindquist, R. A., Moffat, J., et al. (2006) · Genome biology · doi:10.1186/gb-2006-7-10-r100 · Cited by 5,172 · see citing works
Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., Volz, S. E., Joung, J., John van der, O. O. S. T., Regev, A., et al. (2015) · Cell · doi:10.1016/j.cell.2015.09.038 · Cited by 4,750 · see citing works
DNA targeting specificity of RNA-guided Cas9 nucleases
Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann, S., Agarwala, V., Li, Y., Fine, E. J., Wu, X., Shalem, O., et al. (2013) · Nature Biotechnology · doi:10.1038/nbt.2647 · Cited by 4,741 · see citing works
Differential analysis of gene regulation at transcript resolution with RNA-seq
Trapnell, C., Hendrickson, D. G., Sauvageau, M., Goff, L., Rinn, J. L., & Pachter, L. (2013) · Nature Biotechnology · doi:10.1038/nbt.2450 · Cited by 3,492 · see citing works
Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex
Konermann, S., Brigham, M. D., Trevino, A. E., Joung, J., Abudayyeh, O. O., Barcena, C., Hsu, P. D., Habib, N., Gootenberg, J. S., Nishimasu, H., et al. (2015) · Nature · doi:10.1038/nature14136 · Cited by 2,825 · see citing works
In vivo genome editing using Staphylococcus aureus Cas9
Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., et al. (2015) · Nature · doi:10.1038/nature14299 · Cited by 2,687 · see citing works
Crystal structure of Cas9 in complex with guide RNA and target DNA
Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I., Dohmae, N., Ishitani, R., Zhang, F., & Nureki, O. (2014) · Cell · doi:10.1016/j.cell.2014.02.001 · Cited by 2,306 · see citing works
Viral Immune Evasion Due to Persistence of Activated T Cells Without Effector Function
Zajac, A. J., Blattman, J. N., Murali-Krishna, K., Sourdive, D. J. D., Suresh, M., Altman, J. D., & Ahmed, R. (1998) · The Journal of Experimental Medicine · doi:10.1084/jem.188.12.2205 · Cited by 1,978 · see citing works
MiXCR: software for comprehensive adaptive immunity profiling
Bolotin, D. A., Poslavskii, S., Mitrophanov, I., Shugay, M., Mamedov, I. Z., Putintseva, E. V., & Chudakov, D. M. (2015) · Nature Methods · doi:10.1038/nmeth.3364 · Cited by 1,730 · see citing works
Engineered CRISPR-Cas9 nucleases with altered PAM specificities
Kleinstiver, B. P., Prew, M. S., Tsai, S. Q., Topkar, V. V., Nguyen, N. T., Zheng, Z., Gonzales, A. P. W., Li, Z., Peterson, R. T., Yeh, J.-R. J., et al. (2015) · Nature · doi:10.1038/nature14592 · Cited by 1,624 · see citing works
Highly efficient Cas9-mediated transcriptional programming
Chavez, A., Scheiman, J., Vora, S., Pruitt, B. W., Tuttle, M., P R Iyer, E., Lin, S., Kiani, S., Guzman, C. D., Wiegand, D. J., et al. (2015) · Nature Methods · doi:10.1038/nmeth.3312 · Cited by 1,622 · see citing works
Self-citationAnalysis of AAV Serotypes 1–9 Mediated Gene Expression and Tropism in Mice After Systemic Injection
Zincarelli, C., Soltys, S., Rengo, G., & Rabinowitz, J. E. (2008) · Molecular Therapy · doi:10.1038/mt.2008.76 · Cited by 1,490 · see citing works
Zolotukhin, S., Byrne, B. J., Mason, E. G., Zolotukhin, I., Potter, M., Chesnut, K. A., Summerford, C., Samulski, R. J., & Muzyczka, N. (1999) · Gene Therapy · doi:10.1038/sj.gt.3300938 · Cited by 1,317 · see citing works
Structures of Cas9 endonucleases reveal RNA-mediated conformational activation
Jinek, M., Jiang, F., Taylor, D. W., Sternberg, S. H., Kaya, E., Ma, E., Anders, C., Hauer, M., Zhou, K., Lin, S., et al. (2014) · Science · doi:10.1126/science.1247997 · Cited by 1,230 · see citing works
In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy
Nelson, C. E., Hakim, C. H., Ousterout, D. G., Thakore, P. I., Moreb, E. A., Rivera, R. M. C., Madhavan, S., Pan, X., Ran, F. A., Yan, W. X., et al. (2016) · Science · doi:10.1126/science.aad5143 · Cited by 1,144 · see citing works
Clades of Adeno-Associated Viruses Are Widely Disseminated in Human Tissues
Gao, G., Vandenberghe, L. H., Alvira, M. R., Lu, Y., Calcedo, R., Zhou, X., & Wilson, J. M. (2004) · Journal of Virology · doi:10.1128/jvi.78.12.6381-6388.2004 · Cited by 1,070 · see citing works
In vivo gene editing in dystrophic mouse muscle and muscle stem cells
Tabebordbar, M., Zhu, K., Cheng, J. K. W., Chew, W. L., Widrick, J. J., Yan, W. X., Maesner, C., Wu, E. Y., Xiao, R., Ran, F. A., et al. (2016) · Science · doi:10.1126/science.aad5177 · Cited by 1,046 · see citing works
Self-citationElectroporation and RNA interference in the rodent retina in vivo and in vitro
Matsuda, T., & Cepko, C. L. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2235688100 · Cited by 1,032 · see citing works
Boutin, S., Monteilhet, V., Veron, P., Leborgne, C., Benveniste, O., Montus, M. F., & Masurier, C. (2010) · Human Gene Therapy · doi:10.1089/hum.2009.182 · Cited by 970 · see citing works
Gene transfer into muscle by electroporation in vivo
Aihara, H., & Miyazaki, J. (1998) · Nature Biotechnology · doi:10.1038/nbt0998-867 · Cited by 945 · see citing works
Long, C., Amoasii, L., Mireault, A. A., McAnally, J., Li, H. Z., Sanchez‐Ortiz, E., Bhattacharyya, S., Shelton, J. M., Bassel‐Duby, R., & Olson, E. N. (2015) · Science · doi:10.1126/science.aad5725 · Cited by 941 · see citing works
Yin, H., Song, C.-Q., Dorkin, J. R., Zhu, L. J., Li, Y., Wu, Q., Park, A., Yang, J., Suresh, S., Bizhanova, A., et al. (2016) · Nature Biotechnology · doi:10.1038/nbt.3471 · Cited by 876 · see citing works
Pipkin, M. E., Sacks, J. A., Cruz‐Guilloty, F., Lichtenheld, M. G., Bevan, M. J., & Rao, A. (2010) · Immunity · doi:10.1016/j.immuni.2009.11.012 · Cited by 799 · see citing works
Orthogonal Cas9 proteins for RNA-guided gene regulation and editing
Esvelt, K. M., Mali, P., Braff, J. L., Moosburner, M., Yaung, S. J., & Church, G. M. (2013) · Nature Methods · doi:10.1038/nmeth.2681 · Cited by 786 · see citing works
Self-citationIn vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9
Swiech, L., Heidenreich, M., Banerjee, A., Habib, N., Li, Y., Trombetta, J., Sur, M., & Zhang, F. (2015) · Nature Biotechnology · doi:10.1038/nbt.3055 · Cited by 772 · see citing works
A split-Cas9 architecture for inducible genome editing and transcription modulation
Zetsche, B., Volz, S. E., & Zhang, F. (2015) · Nature Biotechnology · doi:10.1038/nbt.3149 · Cited by 704 · see citing works
Dystrophin Immunity in Duchenne's Muscular Dystrophy
Mendell, J. R., Campbell, K., Rodino-Klapac, L., Sahenk, Z., Shilling, C., Lewis, S., Bowles, D., Gray, S., Li, C., Galloway, G., et al. (2010) · New England Journal of Medicine · doi:10.1056/nejmoa1000228 · Cited by 655 · see citing works
Photoactivatable CRISPR-Cas9 for optogenetic genome editing
Nihongaki, Y., Kawano, F., Nakajima, T., & Sato, M. (2015) · Nature Biotechnology · doi:10.1038/nbt.3245 · Cited by 609 · see citing works
Production and characterization of adeno-associated viral vectors
Grieger, J. C., Choi, V. W., & Samulski, R. J. (2006) · Nature Protocols · doi:10.1038/nprot.2006.207 · Cited by 567 · see citing works
A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice
Yang, Y., Wang, L., Bell, P., McMenamin, D., He, Z., White, J., Yu, H., Xu, C., Morizono, H., Musunuru, K., et al. (2016) · Nature Biotechnology · doi:10.1038/nbt.3469 · Cited by 556 · see citing works
Antibody-based protection against HIV infection by vectored immunoprophylaxis
Balazs, A. B., Chen, J., Hong, C. M., Rao, D. S., Yang, L., & Baltimore, D. (2012) · Nature · doi:10.1038/nature10660 · Cited by 545 · see citing works
Signal 3 Determines Tolerance versus Full Activation of Naive CD8 T Cells
Curtsinger, J., Lins, D. C., & Mescher, M. F. (2003) · The Journal of Experimental Medicine · doi:10.1084/jem.20021910 · Cited by 510 · see citing works
Aurnhammer, C., Haase, M., Müther, N., Mück-Häusl, M., Rauschhuber, C., Huber, I., Nitschko, H., Busch, U., Sing, A., Ehrhardt, A., et al. (2012) · Human Gene Therapy Methods · doi:10.1089/hgtb.2011.034 · Cited by 495 · see citing works
Comprehensive serological profiling of human populations using a synthetic human virome
Xu, G. J., Kula, T., Xu, Q., Li, M. Z., Vernon, S. D., Ndung’u, T., Ruxrungtham, K., Sánchez, J., Brander, C., Chung, R. T., et al. (2015) · Science · doi:10.1126/science.aaa0698 · Cited by 478 · see citing works
Biocontainment of genetically modified organisms by synthetic protein design
Mandell, D. J., Lajoie, M. J., Mee, M. T., Takeuchi, R., Kuznetsov, G., Norville, J. E., Gregg, C. J., Stoddard, B. L., & Church, G. M. (2015) · Nature · doi:10.1038/nature14121 · Cited by 474 · see citing works
Self-citationCrystal Structure of Staphylococcus aureus Cas9
Nishimasu, H., Cong, L., Yan, W. X., Ran, F. A., Zetsche, B., Li, Y., Kurabayashi, A., Ishitani, R., Zhang, F., & Nureki, O. (2015) · Cell · doi:10.1016/j.cell.2015.08.007 · Cited by 430 · see citing works
Towards error-free profiling of immune repertoires
Shugay, M., Britanova, O. V., Merzlyak, E. M., Turchaninova, M. A., Mamedov, I. Z., Tuganbaev, T., Bolotin, D. A., Staroverov, D. B., Putintseva, E. V., Plevová, K., et al. (2014) · Nature Methods · doi:10.1038/nmeth.2960 · Cited by 428 · see citing works
Development of an intein-mediated split-Cas9 system for gene therapy
Truong, D.-J. J., Kühner, K., Kühn, R., Werfel, S., Engelhardt, S., Wurst, W., & Ortiz, O. (2015) · Nucleic Acids Research · doi:10.1093/nar/gkv601 · Cited by 371 · see citing works
Structure and Engineering of Francisella novicida Cas9
Hirano, H., Gootenberg, J. S., Horii, T., Abudayyeh, O. O., Kimura, M., Hsu, P. D., Nakane, T., Ishitani, R., Hatada, I., Zhang, F., et al. (2016) · Cell · doi:10.1016/j.cell.2016.01.039 · Cited by 361 · see citing works
Wang, D., Mou, H., Li, S., Li, Y., Hough, S., Tran, K., Li, J., Yin, H., Anderson, D. G., Sontheimer, E. J., et al. (2015) · Human Gene Therapy · doi:10.1089/hum.2015.087 · Cited by 345 · see citing works
Cas9 gRNA engineering for genome editing, activation and repression
Kiani, S., Chavez, A., Tuttle, M. L., Hall, R. N., Chari, R., Ter‐Ovanesyan, D., Qian, J., Pruitt, B. W., Beal, J., Vora, S., et al. (2015) · Nature Methods · doi:10.1038/nmeth.3580 · Cited by 335 · see citing works
Self-citationA robust system for production of minicircle DNA vectors
Kay, M. A., He, C.-Y., & Chen, Z.-Y. (2010) · Nature Biotechnology · doi:10.1038/nbt.1708 · Cited by 329 · see citing works
Rational design of a split-Cas9 enzyme complex
Wright, A. V., Sternberg, S. H., Taylor, D. W., Staahl, B. T., Bardales, J. A., Kornfeld, J. E., & Doudna, J. A. (2015) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1501698112 · Cited by 279 · see citing works
Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease
Dahlman, J. E., Abudayyeh, O. O., Joung, J., Gootenberg, J. S., Zhang, F., & Konermann, S. (2015) · Nature Biotechnology · doi:10.1038/nbt.3390 · Cited by 266 · see citing works
CRISPR/Cas9‐mediated genome engineering: An adeno‐associated viral (AAV) vector toolbox
Senís, E., Fatouros, C., Große, S., Wiedtke, E., Niopek, D., Mueller, A., Börner, K., & Grimm, D. (2014) · Biotechnology Journal · doi:10.1002/biot.201400046 · Cited by 262 · see citing works
Structural insight into the unique properties of adeno-associated virus serotype 9
DiMattia, M. A., Nam, H.-J., Van Vliet, K., Mitchell, M., Bennett, A., Gurda, B. L., McKenna, R., Olson, N. H., Sinkovits, R. S., Potter, M., et al. (2012) · Journal of Virology · doi:10.1128/jvi.07232-11 · Cited by 219 · see citing works
Mamedov, I. Z., Britanova, O. V., Zvyagin, I. V., Turchaninova, M. A., Bolotin, D. A., Putintseva, E. V., Lebedev, Y. B., & Chudakov, D. M. (2013) · Frontiers in Immunology · doi:10.3389/fimmu.2013.00456 · Cited by 172 · see citing works
Adachi, K., Enoki, T., Kawano, Y., Veraz, M., & Nakai, H. (2014) · Nature Communications · doi:10.1038/ncomms4075 · Cited by 149 · see citing works
Gene Transfer into Muscle by Electroporation In Vivo
Miyazaki, J., & Aihara, H. (2003) · Humana Press eBooks · doi:10.1385/1-59259-141-8:049 · Cited by 140 · see citing works
The Complex and Evolving Story of T cell Activation to AAV Vector-encoded Transgene Products
Mays, L. E., & Wilson, J. M. (2010) · Molecular Therapy · doi:10.1038/mt.2010.250 · Cited by 126 · see citing works
Digital cell quantification identifies global immune cell dynamics during influenza infection
Altboum, Z., Steuerman, Y., David, E., Barnett‐Itzhaki, Z., Valadarsky, L., Keren‐Shaul, H., Meningher, T., Mendelson, E., Mandelboim, M., Gat‐Viks, I., et al. (2014) · Molecular Systems Biology · doi:10.1002/msb.134947 · Cited by 119 · see citing works
Protein Trans -Splicing as a Means for Viral Vector-Mediated In Vivo Gene Therapy
Li, J., Sun, W., Wang, B., Xiao, X., & Liu, X. (2008) · Human Gene Therapy · doi:10.1089/hum.2008.009 · Cited by 115 · see citing works
Lin, S.-W., Hensley, S. E., Tatsis, N., Lasaro, M. O., & Ertl, H. C. J. (2007) · Journal of Clinical Investigation · doi:10.1172/jci33138 · Cited by 88 · see citing works
Velazquez, V. M., Bowen, D. G., & Walker, C. M. (2009) · Blood · doi:10.1182/blood-2008-01-131375 · Cited by 46 · see citing works
Fine, E. J., Appleton, C. M., White, D. E., Brown, M. T., Deshmukh, H., Kemp, M. L., & Bao, G. (2015) · Scientific Reports · doi:10.1038/srep10777 · Cited by 41 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Adeno-associated virus vector as a platform for gene therapy delivery
Wang, D., Tai, P. W. L., & Gao, G. (2019) · Nature Reviews Drug Discovery · doi:10.1038/s41573-019-0012-9 · Cited by 2,444 · see citing works
Dunbar, C. E., High, K. A., Joung, J. K., Kohn, D. B., Ozawa, K., & Sadelain, M. (2018) · Science · doi:10.1126/science.aan4672 · Cited by 1,410 · see citing works
Viral vector platforms within the gene therapy landscape
Bulcha, J. T., Wang, Y., Ma, H., Tai, P. W. L., & Gao, G. (2021) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-021-00487-6 · Cited by 1,404 · see citing works
Delivering CRISPR: a review of the challenges and approaches
Lino, C. A., Harper, J. C., Carney, J. P., & Timlin, J. A. (2018) · Drug Delivery · doi:10.1080/10717544.2018.1474964 · Cited by 1,185 · see citing works
Identification of preexisting adaptive immunity to Cas9 proteins in humans
Charlesworth, C. T., Deshpande, P. S., Dever, D. P., Camarena, J., Lemgart, V. T., Cromer, M. K., Vakulskas, C. A., Collingwood, M. A., Zhang, L., Bode, N. M., et al. (2019) · Nature Medicine · doi:10.1038/s41591-018-0326-x · Cited by 1,093 · see citing works
In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni
Kim, E., Koo, T., Park, S. W., Kim, D., Kim, K., Cho, H.-Y., Song, D. W., Lee, K. J., Jung, M. H., Kim, S., et al. (2017) · Nature Communications · doi:10.1038/ncomms14500 · Cited by 715 · see citing works
Advances in the delivery of RNA therapeutics: from concept to clinical reality
Kaczmarek, J. C., Kowalski, P. S., & Anderson, D. G. (2017) · Genome Medicine · doi:10.1186/s13073-017-0450-0 · Cited by 656 · see citing works
Wang, H.-X., Li, M., Lee, C. M., Chakraborty, S., Kim, H.-W., Bao, G., & Leong, K. W. (2017) · Chemical Reviews · doi:10.1021/acs.chemrev.6b00799 · Cited by 560 · see citing works
Structure-guided chemical modification of guide RNA enables potent non-viral in vivo genome editing
Yin, H., Song, C.-Q., Suresh, S., Wu, Q., Walsh, S., Rhym, L. H., Mintzer, E., Bolukbasi, M. F., Zhu, L. J., Kauffman, K., et al. (2017) · Nature Biotechnology · doi:10.1038/nbt.4005 · Cited by 499 · see citing works
In vivo target gene activation via CRISPR/Cas9-mediated trans-epigenetic modulation
Liao, H.-K., Hatanaka, F., Araoka, T., Reddy, P., Wu, M.-Z., Sui, Y., Yamauchi, T., Sakurai, M., O’Keefe, D. D., Núñez-Delicado, E., et al. (2017) · Cell · doi:10.1016/j.cell.2017.10.025 · Cited by 451 · see citing works
High prevalence of Streptococcus pyogenes Cas9-reactive T cells within the adult human population
Wagner, D. L., Amini, L., Wendering, D. J., Burkhardt, L.-M., Akyüz, L., Reinke, P., Volk, H.-D., & Schmueck-Henneresse, M. (2019) · Nature Medicine · doi:10.1038/s41591-018-0204-6 · Cited by 399 · see citing works
Staahl, B. T., Benekareddy, M., Coulon-Bainier, C., Banfal, A. A., Floor, S. N., Sabo, J. K., Urnes, C., Munares, G. A., Ghosh, A., & Doudna, J. A. (2017) · Nature Biotechnology · doi:10.1038/nbt.3806 · Cited by 355 · see citing works
Nrl knockdown by AAV-delivered CRISPR/Cas9 prevents retinal degeneration in mice
Yu, W., Mookherjee, S., Chaitankar, V., Hiriyanna, S., Kim, J.-W., Brooks, M., Ataeijannati, Y., Sun, X., Dong, L., Li, T., et al. (2017) · Nature Communications · doi:10.1038/ncomms14716 · Cited by 271 · see citing works
CRISPR-Cas9 for cancer therapy: Opportunities and challenges
Chen, M., Mao, A., Xu, M., Weng, Q., Mao, J., & Ji, J. (2019) · Cancer Letters · doi:10.1016/j.canlet.2019.01.017 · Cited by 272 · see citing works
Cas9 immunity creates challenges for CRISPR gene editing therapies
Crudele, J. M., & Chamberlain, J. S. (2018) · Nature Communications · doi:10.1038/s41467-018-05843-9 · Cited by 259 · see citing works
Delivery of CRISPR-Cas tools for in vivo genome editing therapy: Trends and challenges
Taha, E. A., Lee, J., & Hotta, A. (2022) · Journal of Controlled Release · doi:10.1016/j.jconrel.2022.01.013 · Cited by 259 · see citing works
Prevalence of Pre-existing Antibodies to CRISPR-Associated Nuclease Cas9 in the USA Population
Simhadri, V. L., McGill, J., McMahon, S., Wang, J., Jiang, H., & Sauna, Z. E. (2018) · Molecular Therapy - Methods & Clinical Development · doi:10.1016/j.omtm.2018.06.006 · Cited by 241 · see citing works
Methods and Applications of CRISPR-Mediated Base Editing in Eukaryotic Genomes
Hess, G. T., Tycko, J., Yao, D., & Bassik, M. C. (2017) · Molecular Cell · doi:10.1016/j.molcel.2017.09.029 · Cited by 234 · see citing works
AAV-CRISPR Gene Editing Is Negated by Pre-existing Immunity to Cas9
Li, A., Tanner, M. R., Lee, C. M., Hurley, A. E., De Giorgi, M., Jarrett, K. E., Davis, T. H., Doerfler, A. M., Bao, G., Beeton, C., et al. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.04.017 · Cited by 230 · see citing works
Multifunctional CRISPR-Cas9 with Engineered Immunosilenced Human T Cell Epitopes
Ferdosi, S. R., Ewaisha, R., Moghadam, F., Krishna, S., Park, J. G., Ebrahimkhani, M. R., Kiani, S., & Anderson, K. S. (2019) · Nature Communications · doi:10.1038/s41467-019-09693-x · Cited by 204 · see citing works
Hakim, C. H., Kumar, S. R. P., Pérez-López, D. O., Wasala, N. B., Zhang, D., Yue, Y., Teixeira, J., Pan, X., Zhang, K., Million, E. D., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-26830-7 · Cited by 175 · see citing works
Evaluating and enhancing target specificity of gene-editing nucleases and deaminases
Kim, D., Luk, K., Wolfe, S. A., & Kim, J.-S. (2019) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-013118-111730 · Cited by 171 · see citing works
Adeno-associated viral vector-mediated immune responses: Understanding barriers to gene delivery
Nidetz, N. F., McGee, M. C., Tse, L. V., Li, C., Cong, L., Li, Y., & Huang, W. (2020) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2019.107453 · Cited by 174 · see citing works
Dual-AAV delivering split prime editor system for in vivo genome editing
Zhi, S., Chen, Y., Wu, G., Wen, J., Wu, J., Liu, Q., Li, Y., Kang, R., Hu, S., Wang, J., et al. (2022) · Molecular Therapy · doi:10.1016/j.ymthe.2021.07.011 · Cited by 174 · see citing works
Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease for Cell-Type-Specific Gene Editing
Rouet, R., Thuma, B. A., Roy, M. D., Lintner, N. G., Rubitski, D. M., Finley, J. E., Wisniewska, H. M., Mendonsa, R., Hirsh, A., de Oñate, L., et al. (2018) · Journal of the American Chemical Society · doi:10.1021/jacs.8b01551 · Cited by 163 · see citing works
Lau, C.-H., & Suh, Y. (2017) · F1000Research · doi:10.12688/f1000research.11243.1 · Cited by 161 · see citing works
In Situ Gene Therapy via AAV-CRISPR-Cas9-Mediated Targeted Gene Regulation
Moreno, A. M., Fu, X., Zhu, J., Katrekar, D., Shih, Y.-R. V., Marlett, J., Cabotaje, J., Tat, J., Naughton, J., Lisowski, L., et al. (2018) · Molecular Therapy · doi:10.1016/j.ymthe.2018.04.017 · Cited by 159 · see citing works
Self-citationVSV-G-Enveloped Vesicles for Traceless Delivery of CRISPR-Cas9
Montagna, C., Petris, G., Casini, A., Maule, G., Franceschini, G. M., Zanella, I., Conti, L., Arnoldi, F., Burrone, O. R., Zentilin, L., et al. (2018) · Molecular Therapy Nucleic Acids · doi:10.1016/j.omtn.2018.05.010 · Cited by 117 · see citing works
Seitz, I. P., Michalakis, S., Wilhelm, B., Reichel, F. F., Ochakovski, G. A., Zrenner, E., Ueffing, M., Biel, M., Wissinger, B., Bartz-Schmidt, K. U., et al. (2017) · Investigative Opthalmology & Visual Science · doi:10.1167/iovs.17-22473 · Cited by 108 · see citing works
Gaj, T., Staahl, B. T., Rodrigues, G. M. C., Limsirichai, P., Ekman, F. K., Doudna, J. A., & Schaffer, D. V. (2017) · Nucleic Acids Research · doi:10.1093/nar/gkx154 · Cited by 89 · see citing works
Sanz, D. J., Hollywood, J. A., Scallan, M. F., & Harrison, P. T. (2017) · PLOS ONE · doi:10.1371/journal.pone.0184009 · Cited by 92 · see citing works
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 · see citing works
Gene Therapy Intervention in Neovascular Eye Disease: A Recent Update
Lin, F.-L., Wang, P.-Y., Chuang, Y.-F., Wang, J.-H., Wong, V. H. Y., Bui, B. V., & Liu, G.-S. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.06.029 · Cited by 74 · see citing works
Zinsli, L. V., Stierlin, N., Loessner, M. J., & Schmelcher, M. (2021) · Computational and Structural Biotechnology Journal · doi:10.1016/j.csbj.2020.12.024 · Cited by 63 · see citing works
Cas9-directed immune tolerance in humans—a model to evaluate regulatory T cells in gene therapy?
Wagner, D. L., Peter, L., & Schmueck-Henneresse, M. (2021) · Gene Therapy · doi:10.1038/s41434-021-00232-2 · Cited by 63 · see citing works
Oligonucleotide conjugated multi-functional adeno-associated viruses
Katrekar, D., Moreno, A. M., Chen, G., Worlikar, A., & Mali, P. (2018) · Scientific Reports · doi:10.1038/s41598-018-21742-x · Cited by 57 · see citing works
Self-citationRen, D., Fisson, S., Dalkara, D., & Ail, D. (2022) · Pharmaceutics · doi:10.3390/pharmaceutics14091973 · Cited by 52 · see citing works
Tariq, A., Mushtaq, M., Yaqoob, H., Bhat, B. A., Zargar, S. M., Raza, A., Ali, S., Charagh, S., Mubarik, M. S., Zaman, Q. U., et al. (2023) · GM Crops & Food · doi:10.1080/21645698.2023.2219111 · Cited by 45 · see citing works
Rational design of a compact CRISPR-Cas9 activator for AAV-mediated delivery
Vora, S., Cheng, J., Xiao, R., VanDusen, N. J., Quintino, L., Pu, W. T., Vandenberghe, L. H., Chavez, A., & Church, G. (2018) · doi:10.1101/298620 · Cited by 42 · see citing works
Self-citationLohia, A., Sahel, D. K., Salman, M., Singh, V., Mariappan, I., Mittal, A., & Chitkara, D. (2022) · Asian Journal of Pharmaceutical Sciences · doi:10.1016/j.ajps.2022.02.001 · Cited by 33 · see citing works
Design 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 · see citing works
TLR9-independent CD8+ T cell responses in hepatic AAV gene transfer through IL-1R1-MyD88 signaling
Kumar, S. R. P., Biswas, M., Cao, D., Arisa, S., Muñoz-Melero, M., Lam, A. K., Piñeros, A. R., Kapur, R., Kaisho, T., Kaufman, R. J., et al. (2024) · Molecular Therapy · doi:10.1016/j.ymthe.2023.11.029 · Cited by 31 · see citing works
Split AAV8 Mediated ABCA4 Expression for Gene Therapy of Mouse Stargardt Disease (STGD1)
Li, R., Jing, Q., She, K., Wang, Q., Jin, X., Zhao, Q., Su, J., Song, L., Fu, J., Wu, X., et al. (2023) · Human Gene Therapy · doi:10.1089/hum.2023.017 · Cited by 21 · see citing works
Evasion of Pre-Existing Immunity to Cas9: A Prerequisite for Successful Genome Editing In Vivo?
Wignakumar, T., & Fairchild, P. J. (2019) · Current Transplantation Reports · doi:10.1007/s40472-019-00237-2 · Cited by 19 · see citing works
- Authors
- Chew, W. L., Tabebordbar, M., Cheng, J. K. W., Mali, P., Wu, E. Y., Ng, A. H. M., Zhu, K., Wagers, A. J., & Church, G. M.
- Affiliations
- Harvard University (US)
- Corresponding author
- Wagers, Amy J
- Church, George M
- ORCID
- Venue
- Nature Methods
- Published
- 2016-09-05
- Volume
- 13
- Issue
- 10
- Pages
- 868-874
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC5374744
- OpenAlex
- W2508903105
- Cited by
- 617
- References
- 63
- Citations by year
- 2026: 28 · 2025: 58 · 2024: 45 · 2023: 65 · 2022: 78 · 2021: 84 · 2020: 66 · 2019: 73 · 2018: 56 · 2017: 57 · 2016: 6
- FWCI
- 33.27
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (acceptedVersion)
- Funders
- Howard Hughes Medical Institute
- National Heart, Lung, and Blood Institute (U01HL100402)
- National Human Genome Research Institute (P50HG005550)
- National Eye Institute (PN2EY018244)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- 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
- Keywords
- adeno-associated virus vectors, CRISPR/Cas9 genome editing, host immune response, antigenicity, postnatal mice, transcriptional regulation, cell damage
- MeSH terms
- Gene Editing, Dependovirus, Animals, Enzyme-Linked Immunosorbent Assay, Genetic Engineering, Genetic Vectors, Humans, Mice, Inbred C57BL, Gene Transfer Techniques, Reverse Transcriptase Polymerase Chain Reaction, Mice, HEK293 Cells, CRISPR-Cas Systems
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
