Cited by 230CRISPR and Genetic EngineeringVirus-based gene therapy researchRNA Interference and Gene Delivery

The DOI matches its Crossref record; 39 of the 40 references it cites were found in a registry or an index.

AAV-CRISPR Gene Editing Is Negated by Pre-existing Immunity to Cas9

Open access

Li, Ang, Tanner, Mark R., Lee, Ciaran M., Hurley, Ayrea E., De Giorgi, Marco, Jarrett, Kelsey E., Davis, Timothy H., Doerfler, Alexandria M., Bao, Gang, Beeton, Christine, Lagor, William R.·2020·Molecular Therapy

DOI 10.1016/j.ymthe.2020.04.017PMC PMC7264438

Open this paper in CoChat — free account Open PDF

Abstract

Adeno-associated viral (AAV) vectors are a leading candidate for the delivery of CRISPR-Cas9 for therapeutic genome editing in vivo . However, AAV-based delivery involves persistent expression of the Cas9 nuclease, a bacterial protein. Recent studies indicate a high prevalence of neutralizing antibodies and T cells specific to the commonly used Cas9 orthologs from Streptococcus pyogenes (SpCas9) and Staphylococcus aureus (SaCas9) in humans. We tested in a mouse model whether pre-existing immunity to SaCas9 would pose a barrier to liver genome editing with AAV packaging CRISPR-Cas9. Although efficient genome editing occurred in mouse liver with pre-existing SaCas9 immunity, this was accompanied by an increased proportion of CD8 + T cells in the liver. This cytotoxic T cell response was characterized by hepatocyte apoptosis, loss of recombinant AAV genomes, and complete elimination of genome-edited cells, and was followed by compensatory liver regeneration. Our results raise important efficacy and safety concerns for CRISPR-Cas9-based in vivo genome editing in the liver.

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

Crossref · 10.1016/j.ymthe.2020.04.017

Cites

39 of 39 references verified0 citation errors · 0 not checkable

Statistics

Graphical abstract CC BY-NC-ND · PubMed Central

Figure 1 Induction of Immune Memory against SaCas9 in C57BL/6 Mice (A) Mice were initially immunized with either purified ovalbumin (Ova) or SaCas9 protein. One week after immunization, mice were challenged with either ovalbumin or SaCas9, and ear thickness was measured after 24 h. (B) Histological analysis of the left ear of mice in (A) stained with hematoxylin and eosin. ∗p < 0.05, ∗∗p < 0.01; ns, not significant. Scale bar, 50 μm. (C) Representative H&E staining of mouse ears.CC BY-NC-ND · PubMed Central

Figure 2 AAV-CRISPR Gene Editing of Mice Immunized against SaCas9 Elicits a Robust CD8+ T Cell Response (A) Mice were immunized with either ovalbumin or SaCas9 protein 1 week prior to co-delivery of AAV vectors encoding a gRNA targeting Ldlr with SaCas9 and GFP. Tissues were harvested at 1, 2, 4, 6, and 12 weeks post-injection. (B) CD8+ T cells as a percentage of total lymphocytes in the liver were measured by flow cytometry. (C) CD8 mRNA expression between mice immunized against ovalbumin and SaCas9 measured by qPCR. (D) Representative TUNEL staining in mouse liver sections. (E) Serum alanine transaminase (ALT) activity levels indicating liver damage. (F) Representative Ki-67 staining of mouse liver sections. (G) Ki-67 mRNA expression in mouse liver. Group sizes in (C), (E), and (G) are n = 7, except for ovalbumin 6 weeks (n = 6) and SaCas9 1 week (n = 6).CC BY-NC-ND · PubMed Central

Figure 3 Pre-existing Immunity to SaCas9 Results in Elimination of Gene-Edited Hepatocytes following AAV Transduction (A) AAV genome copies of GFP transgene measured via qPCR. (B) Fold change of GFP genome copies relative to ovalbumin-treated mice. (C) AAV genome copies of SaCas9 transgene measured via qPCR. (D) Fold change of SaCas9 genome copies relative to ovalbumin-treated mice. (E) Representative western blots of LDLR, SaCas9 (hemagglutinin, HA-Tag), and GFP in mouse liver. Asterisk indicates non-specific band by the HA-Tag antibody. (F) Ldlr indels in mouse liver samples were measured via deep sequencing.CC BY-NC-ND · PubMed Central

Figure 4 CD8+ T Cell and Treg Responses in SaCas9-Immunized Mice after Treatment with Either AAV-GFP or AAV-CRISPR (A) Mice were immunized with SaCas9 protein 1 week prior to delivery of an AAV vector encoding a gRNA targeting Ldlr with SaCas9 or GFP. Tissues were harvested at 2 weeks post-injection. (B) T cells as a percentage of total lymphocytes in the liver were measured via flow cytometry. (C) CD8+ T cells as a percentage of T cells. (D) Percentage of memory CD8+ to total CD8+ T cells. (E) Percentage of activated CD8+ T cells to total CD8+ T cell population. (F) Percentage of activated memory CD8+ T cells to total CD8+ T cell population. (G) Tregs as a percentage of total lymphocytes in the liver measured via flow cytometry. n = 6 per condition.CC BY-NC-ND · PubMed Central

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?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method
Livak, K. J., & Schmittgen, T. D. (2001) · Methods · doi:10.1006/meth.2001.1262 · Cited by 180,860
Mutation of the mouse klotho gene leads to a syndrome resembling ageing
Kuro-o, M., Matsumura, Y., Aizawa, H., Kawaguchi, H., Suga, T., Utsugi, T., Ohyama, Y., Kurabayashi, M., Kaname, T., Kume, E., et al. (1997) · Nature · doi:10.1038/36285 · Cited by 3,837
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
Production of High-Titer Recombinant Adeno-Associated Virus Vectors in the Absence of Helper Adenovirus
Xiao, X., Li, J., & Samulski, R. J. (1998) · Journal of Virology · doi:10.1128/jvi.72.3.2224-2232.1998 · Cited by 1,381
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
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
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
Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy
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 973
Epitope spreading in immune-mediated diseases: implications for immunotherapy
Vanderlugt, C. L., & Miller, S. D. (2002) · Nature reviews. Immunology · doi:10.1038/nri724 · Cited by 947
A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing
Finn, J. D., Smith, A. R., Patel, M. C., Shaw, L., Youniss, M. R., van Heteren, J., Dirstine, T., Ciullo, C., Lescarbeau, R., Seitzer, J., et al. (2018) · Cell Reports · doi:10.1016/j.celrep.2018.02.014 · Cited by 878

Show all 40 references

Sorted by citations

Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy
Nelson, C. E., Wu, Y., Gemberling, M. P., Oliver, M. L., Waller, M. A., Bohning, J. D., Robinson-Hamm, J. N., Bulaklak, K., Rivera, R. M. C., Collier, J. H., et al. (2019) · Nature Medicine · doi:10.1038/s41591-019-0344-3 · Cited by 444
Immunogenicity of CAR T cells in cancer therapy
Wagner, D. L., Fritsche, E., Pulsipher, M. A., Ahmed, N. M., Hamieh, M., Hegde, M. G., Ruella, M., Savoldo, B., Shah, N. N., Turtle, C. J., et al. (2021) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-021-00476-2 · Cited by 318
Low immunogenicity of LNP allows repeated administrations of CRISPR-Cas9 mRNA into skeletal muscle in mice
Kenjo, E., Hozumi, H., Makita, Y., Iwabuchi, K. A., Fujimoto, N., Matsumoto, S., Kimura, M., Amano, Y., Ifuku, M., Naoe, Y., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-26714-w · Cited by 313
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
Microneedle-assisted genome editing: A transdermal strategy of targeting NLRP3 by CRISPR-Cas9 for synergistic therapy of inflammatory skin disorders
Wan, T. Y., Pan, Q., & Yuan, P. (2021) · Science Advances · doi:10.1126/sciadv.abe2888 · Cited by 226
Cas9-specific immune responses compromise local and systemic AAV CRISPR therapy in multiple dystrophic canine models
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
Drug delivery systems for CRISPR-based genome editors
Madigan, V. J., Zhang, F., & Dahlman, J. E. (2023) · Nature Reviews Drug Discovery · doi:10.1038/s41573-023-00762-x · Cited by 193
Covalently closed circular DNA: The ultimate therapeutic target for curing HBV infections
Martínez, M. G., Boyd, A. C., Combe, E., Testoni, B., & Zoulim, F. (2021) · Journal of Hepatology · doi:10.1016/j.jhep.2021.05.013 · Cited by 172
AAV-delivered suppressor tRNA overcomes a nonsense mutation in mice
Wang, J., Zhang, Y., Mendonca, C. A., Yukselen, O., Muneeruddin, K., Ren, L., Liang, J., Zhou, C., Xie, J., Li, J., et al. (2022) · Nature · doi:10.1038/s41586-022-04533-3 · Cited by 144
Transgenic mice for in vivo epigenome editing with CRISPR-based systems
Gemberling, M. P., Siklenka, K., Rodriguez, E., Eisinger, K. R. T., Barrera, A., Liu, F., Kantor, A., Li, L., Cigliola, V., Hazlett, M. F., et al. (2021) · Nature Methods · doi:10.1038/s41592-021-01207-2 · Cited by 155
Immunogenicity of CRISPR therapeutics—Critical considerations for clinical translation
Ewaisha, R., & Anderson, K. S. (2023) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2023.1138596 · Cited by 153
Viral 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
CRISPR/Cas9 systems: Delivery technologies and biomedical applications
Du, Y., Liu, Y., Hu, J., Peng, X., & Liu, Z. (2023) · Asian Journal of Pharmaceutical Sciences · doi:10.1016/j.ajps.2023.100854 · Cited by 140
Immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy
Moreno, A. M., Palmer, N. D., Alemán, F., Chen, G., Pla, A., Jiang, N. J., Chew, W. L., Law, M., & Mali, P. (2019) · Nature Biomedical Engineering · doi:10.1038/s41551-019-0431-2 · Cited by 132
The NIH Somatic Cell Genome Editing program
Saha, K., Sontheimer, E. J., Brooks, P. J., Dwinell, M. R., Gersbach, C. A., Liu, D. R., Murray, S. A., Tsai, S. Q., Wilson, R. C., Anderson, D. G., et al. (2021) · Nature · doi:10.1038/s41586-021-03191-1 · Cited by 127
Self-citation
Clinical delivery of circular RNA: Lessons learned from RNA drug development
Young, T. L., Wang, K. C., Varley, A., & Li, B. (2023) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2023.114826 · Cited by 130
Tools for experimental and computational analyses of off-target editing by programmable nucleases
Bao, X. R., Pan, Y., Lee, C. M., Davis, T. H., & Bao, G. (2021) · Nature Protocols · doi:10.1038/s41596-020-00431-y · Cited by 106
Self-citation
CRISPR/Cas9 gene editing for curing sickle cell disease
Park, S. H., & Bao, G. (2021) · Transfusion and Apheresis Science · doi:10.1016/j.transci.2021.103060 · Cited by 118
Self-citation
Ready for Repair? Gene Editing Enters the Clinic for the Treatment of Human Disease
Ernst, M. P. T., Broeders, M., Herrero-Hernandez, P., Oussoren, E., van der Ploeg, A. T., & Pijnappel, W. W. M. P. (2020) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2020.06.022 · Cited by 103
CRISPR-Cas9 gene editing of hepatitis B virus in chronically infected humanized mice
Stone, D., Long, K. R., Loprieno, M. A., Feelixge, H. S. D. S., Kenkel, E. J., Liley, R. M., Rapp, S., Roychoudhury, P., Nguyen, T., Stensland, L., et al. (2020) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2020.11.014 · Cited by 98
Self-citation
Application of CRISPR/Cas9 Technology in Cancer Treatment: A Future Direction
Rabaan, A. A., AlSaihati, H., Bukhamsin, R., Bakhrebah, M. A., Nassar, M. S., Alsaleh, A. A., Alhashem, Y. N., Bukhamseen, A. Y., Al-Ruhimy, K., Alotaibi, M. H., et al. (2023) · Current Oncology · doi:10.3390/curroncol30020152 · Cited by 89
Targeted replacement of full-length CFTR in human airway stem cells by CRISPR-Cas9 for pan-mutation correction in the endogenous locus
Vaidyanathan, S., Baik, R., Chen, L., Bravo, D. T., Suarez, C. J., Abazari, S. M., Salahudeen, A. A., Dudek, A. M., Teran, C. A., Davis, T., et al. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.03.023 · Cited by 70
Self-citation
Comparative analysis of lipid Nanoparticle-Mediated delivery of CRISPR-Cas9 RNP versus mRNA/sgRNA for gene editing in vitro and in vivo
Walther, J., Porenta, D., Wilbie, D., Seinen, C. W., Benne, N., Yang, Q., de Jong, O. G., Lei, Z., & Mastrobattista, E. (2024) · European Journal of Pharmaceutics and Biopharmaceutics · doi:10.1016/j.ejpb.2024.114207 · Cited by 80
Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy
Pickar‐Oliver, A., Gough, V., Bohning, J. D., Liu, S., Robinson-Hamm, J. N., Daniels, H., Majoros, W. H., Devlin, G. W., Asokan, A., & Gersbach, C. A. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.09.003 · Cited by 78
Immunological barriers to haematopoietic stem cell gene therapy
Charlesworth, C. T., Hsu, I., Wilkinson, A. C., & Nakauchi, H. (2022) · Nature reviews. Immunology · doi:10.1038/s41577-022-00698-0 · Cited by 71
Precision RNA base editing with engineered and endogenous effectors
Pfeiffer, L. S., & Stafforst, T. (2023) · Nature Biotechnology · doi:10.1038/s41587-023-01927-0 · Cited by 68
Is Viral Vector Gene Delivery More Effective Using Biomaterials?
Wang, Y., Bruggeman, K. F., Franks, S. J., Gautam, V., Hodgetts, S. I., Harvey, A. R., Williams, R. J., & Nisbet, D. R. (2020) · Advanced Healthcare Materials · doi:10.1002/adhm.202001238 · Cited by 69
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
In utero adenine base editing corrects multi-organ pathology in a lethal lysosomal storage disease
Bose, S. K., White, B. M., Kashyap, M. V., Dave, A., De Bie, F. R., Li, H., Singh, K., Menon, P., Wang, T., Teerdhala, S., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-24443-8 · Cited by 67
Delivery Approaches for Therapeutic Genome Editing and Challenges
Ates, I., Rathbone, T., Stuart, C., Bridges, P. H., & Cottle, R. N. (2020) · Genes · doi:10.3390/genes11101113 · Cited by 63
Immunomodulation in Administration of RAAV: Preclinical and Clinical Adjuvant Pharmacotherapies
Chu, W. S., & Ng, J. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.658038 · Cited by 59
Hematopoietic Stem Cell Gene-Addition/Editing Therapy in Sickle Cell Disease
Germino-Watnick, P., Hinds, M., Le, A., Chu, R., Liu, X., & Uchida, N. (2022) · Cells · doi:10.3390/cells11111843 · Cited by 63
Rational engineering of minimally immunogenic nucleases for gene therapy
Raghavan, R. S., Friedrich, M., King, C., Vo, C.-D.-T., Strebinger, D., Lash, B., Kilian, M., Platten, M., Macrae, R. K., Song, Y., et al. (2025) · Nature Communications · doi:10.1038/s41467-024-55522-1 · Cited by 56
Epigenetic basis and targeting of cancer metastasis
Banerjee, R., Smith, J., Eccles, M. R., Weeks, R. J., & Chatterjee, A. (2021) · Trends in cancer · doi:10.1016/j.trecan.2021.11.008 · Cited by 51
PCSK9 in metabolism and diseases
Ajoolabady, A., Pratico, D., Mazidi, M., Davies, I. G., Lip, G. Y. H., Seidah, N., Libby, P., Kroemer, G., & Ren, J. (2025) · Metabolism · doi:10.1016/j.metabol.2024.156064 · Cited by 49
Genome editing in the mouse brain with minimally immunogenic Cas9 RNPs
Stahl, E. C., Sabo, J. K., Kang, M. H., Allen, R., Applegate, E., Kim, S., Kwon, Y., Seth, A., Lemus, N., Salinas-Rios, V., et al. (2023) · Molecular Therapy · doi:10.1016/j.ymthe.2023.06.019 · Cited by 51
Mini-dCas13X–mediated RNA editing restores dystrophin expression in a humanized mouse model of Duchenne muscular dystrophy
Li, G., Jin, M., Li, Z., Xiao, Q., Lin, J., Yang, D., Liu, Y., Wang, X., Xie, L., Ying, W., et al. (2023) · Journal of Clinical Investigation · doi:10.1172/jci162809 · Cited by 44
Emerging applications of gene editing technologies for the development of climate-resilient crops
Chavhan, R. L., Jaybhaye, S. G., Hinge, V. R., Deshmukh, A. S., Shaikh, U. S., Jadhav, P. K., Kadam, U. S., & Hong, J. C. (2025) · Frontiers in Genome Editing · doi:10.3389/fgeed.2025.1524767 · Cited by 55
“CRISPR/Cas functions as an adaptive immune defense in bacteria, safeguarding their genomic DNA from viral (phage) attacks by degrading the DNA with the help of RNA guidance and the Cas9 protein (Mallapaty, 2019; Li et al., 2020a).”
Liver‐directed gene therapy for inherited metabolic diseases
Baruteau, J., Brunetti‐Pierri, N., & Gissen, P. (2024) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12709 · Cited by 46
Translating CRISPR-Cas Therapeutics: Approaches and Challenges
Tay, L. S., Palmer, N. D., Panwala, R., Chew, W. L., & Mali, P. (2020) · The CRISPR Journal · doi:10.1089/crispr.2020.0025 · Cited by 43
Current strategies employed in the manipulation of gene expression for clinical purposes
Tsai, H., Pietrobon, V., Peng, M., Wang, S., Zhao, L., Marincola, F. M., & Cai, Q. (2022) · Journal of Translational Medicine · doi:10.1186/s12967-022-03747-3 · Cited by 48
Deleting in vivo β-catenin degradation domain in mouse hepatocytes drives hepatocellular carcinoma or hepatoblastoma-like tumors
Loesch, R., Caruso, S., Paradis, V., Godard, C., Gougèlet, A., Renault, G., Picard, S., Tanaka, I., Renoux-Martin, Y., Perret, C., et al. (2021) · Journal of Hepatology · doi:10.1016/j.jhep.2022.02.023 · Cited by 46
Novel therapeutic strategies for chronic hepatitis B
Phillips, S., Jagatia, R., & Chokshi, S. (2022) · Virulence · doi:10.1080/21505594.2022.2093444 · Cited by 44
Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
He, X., Zhang, Z., Xue, J., Wang, Y., Zhang, S., Wei, J., Zhang, C., Wang, J., Urip, B. A., Ngan, C. C., et al. (2022) · Nature Communications · doi:10.1038/s41467-022-34898-y · Cited by 43
Immunologically effective poly(D-lactic acid) nanoparticle enhances anticancer immune response
Xu, W., Su, Y., Ma, Y., Wei, Q., Yang, J.-Z., Zhuang, X., Ding, J., & Chen, X. (2023) · Science China Chemistry · doi:10.1007/s11426-022-1441-7 · Cited by 40
Understanding and overcoming adverse consequences of genome editing on hematopoietic stem and progenitor cells
Lee, B., Lozano, R. J., & Dunbar, C. E. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.09.001 · Cited by 39
Duchenne Muscular Dystrophy Gene Therapy in 2023: Status, Perspective, and Beyond
Duan, D. (2023) · Human Gene Therapy · doi:10.1089/hum.2023.29242.ddu · Cited by 44
Understanding and Tackling Immune Responses to Adeno-Associated Viral Vectors
Verdera, H. C., Unzu, C., Valeri, E., Adriouch, S., González‐Aseguinolaza, G., Mingozzi, F., & Kajaste‐Rudnitski, A. (2023) · Human Gene Therapy · doi:10.1089/hum.2023.119 · Cited by 45
Building CRISPR Gene Therapies for the Central Nervous System
Salomonsson, S. E., & Clelland, C. D. (2024) · JAMA Neurology · doi:10.1001/jamaneurol.2023.4983 · Cited by 42
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
Reprogramming the anti-tumor immune response via CRISPR genetic and epigenetic editing
Alves, E., Taifour, S., Dolcetti, R., Chee, J., Nowak, A. K., Gaudieri, S., & Blancafort, P. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.04.009 · Cited by 40
Repurposing CRISPR-Cas13 systems for robust mRNA trans-splicing
Fiflis, D. N., Rey, N. A., Venugopal-Lavanya, H., Sewell, B., Mitchell-Dick, A., Clements, K. N., Milo, S., Benkert, A. R., Rosales, A., Fergione, S., et al. (2024) · Nature Communications · doi:10.1038/s41467-024-46172-4 · Cited by 36
Development of an in vivo delivery system for CRISPR/Cas9-mediated targeting of hepatitis B virus cccDNA
Kayesh, M. E. H., Amako, Y., Hashem, M. A., Murakami, S., Ogawa, S., Yamamoto, N., Hifumi, T., Miyoshi, N., Sugiyama, M., Tanaka, Y., et al. (2020) · Virus Research · doi:10.1016/j.virusres.2020.198191 · Cited by 39
Molecular Therapies for Myotonic Dystrophy Type 1: From Small Drugs to Gene Editing
Izzo, M., Battistini, J. I., Provenzano, C., Martelli, F., Cardinali, B., & Falcone, G. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23094622 · 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
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
Gene Editing Technologies to Target HBV cccDNA
Martínez, M. G., Smekalova, E. M., Combe, E., Gregoire, F. M., Zoulim, F., & Testoni, B. (2022) · Viruses · doi:10.3390/v14122654 · Cited by 37
Gene therapy for genetic diseases: challenges and future directions
Beibei, Q. I. E., Tuo, J., Chen, F., Ding, H., & Lyu, L. (2025) · MedComm · doi:10.1002/mco2.70091 · Cited by 38
Immune-based therapies in cardiovascular and metabolic diseases: past, present and future
Murphy, A. J., & Febbraio, M. A. (2021) · Nature reviews. Immunology · doi:10.1038/s41577-021-00580-5 · Cited by 35
Liver directed adeno‐associated viral vectors to treat metabolic disease
Chuecos, M. A., & Lagor, W. R. (2023) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.12637 · Cited by 34
Self-citation
RNA lipid nanoparticles as efficient in vivo CRISPR-Cas9 gene editing tool for therapeutic target validation in glioblastoma cancer stem cells
Rouatbi, N., Walters, A. A., Costa, P. M., Qin, Y., Liam‐Or, R., Grant, V., Pollard, S. M., Wang, J. T.-W., & Al‐Jamal, K. T. (2024) · Journal of Controlled Release · doi:10.1016/j.jconrel.2024.09.019 · Cited by 36
Advancements in CRISPR/Cas systems for disease treatment
Xu, Y., Le, H., Wu, Q., Wang, N., & Gong, C. (2025) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2025.05.007 · Cited by 35
Depleting hepatitis B virus relaxed circular DNA is necessary for resolution of infection by CRISPR-Cas9
Kostyushev, D. S., Kostyusheva, A., Brezgin, S., Ponomareva, N. I., Zakirova, N. F., Egorshina, A. Y., Yanvarev, D. V., Олеговна, Б. Е., Sudina, A., Goptar, I. A., et al. (2023) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2023.02.001 · Cited by 34
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
Gene editing for dyslipidemias: New tools to “cut” lipids
Stankov, S. G., & Cuchel, M. (2023) · Atherosclerosis · doi:10.1016/j.atherosclerosis.2023.01.010 · Cited by 34
2021 White Paper on Recent Issues in Bioanalysis: Mass Spec of Proteins, Extracellular Vesicles, CRISPR, Chiral Assays, Oligos; Nanomedicines Bioanalysis; ICH M10 Section 7.1; Non-Liquid & Rare Matrices; Regulatory Inputs ( Part 1A – Recommendations on Endogenous Compounds, Small Molecules, Complex Methods, Regulated Mass Spec of Large Molecules, Small Molecule, PoC & Part 1B - Regulatory Agencies' Inputs on Bioanalysis, Biomarkers, Immunogenicity, Gene & Cell Therapy and Vaccine)
Kaur, S., Alley, S. C., Szapacs, M., Wilson, A., Ciccimaro, E., Su, D., Henderson, N., Chen, L., Garofolo, F., Hengel, S. M., et al. (2022) · Bioanalysis · doi:10.4155/bio-2022-0078 · Cited by 32
Pre-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
Robust genome and cell engineering via in vitro and in situ circularized RNAs
Tong, M., Palmer, N. D., Dailamy, A., Kumar, A., Khaliq, H., Han, S., Finburgh, E. N., Wing, M. K., Hong, C., Xiang, Y., et al. (2024) · Nature Biomedical Engineering · doi:10.1038/s41551-024-01245-z · Cited by 32
Progress in Gene Editing Tools and Their Potential for Correcting Mutations Underlying Hearing and Vision Loss
Botto, C., Dalkara, D., & El-Amraoui, A. (2021) · Frontiers in Genome Editing · doi:10.3389/fgeed.2021.737632 · Cited by 30
Investigation of Cas9 Antibodies in the Human Eye
Toral, M. A., Charlesworth, C. T., Ng, B., Chemudupati, T., Homma, S., Nakauchi, H., Bassuk, A. G., Porteus, M. H., & Mahajan, V. B. (2022) · Nature Communications · doi:10.1038/s41467-022-28674-1 · Cited by 28
Integrated epigenetic and genetic programming of primary human T cells
Goudy, L., Ha, A. S., Borah, A. A., Umhoefer, J. M., Chow, L., Tran, C., Winters, A., Talbot, A., Hernandez, R., Li, Z., et al. (2025) · Nature Biotechnology · doi:10.1038/s41587-025-02856-w · Cited by 38
Synthetic transcription factor engineering for cell and gene therapy
Bhatt, B., García-Díaz, P., & Wink, G. (2023) · Trends in biotechnology · doi:10.1016/j.tibtech.2023.09.010 · Cited by 28
Peeling back the layers of immunogenicity in Cas9-based genomic medicine
Stigzelius, V., Cavallo, A. L., Chandode, R. K., & Nitsch, R. (2025) · Molecular Therapy · doi:10.1016/j.ymthe.2025.06.045 · Cited by 30
Cas9-derived peptides presented by MHC Class II that elicit proliferation of CD4+ T-cells
Simhadri, V. L., Hopkins, L., McGill, J. R., Duke, B. R., Mukherjee, S., Zhang, K., & Sauna, Z. E. (2021) · Nature Communications · doi:10.1038/s41467-021-25414-9 · Cited by 29
A universal strategy for AAV delivery of base editors to correct genetic point mutations in neonatal PKU mice
Zhou, 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
Nonviral Delivery of CRISPR/Cas Systems in mRNA Format
Liu, Z., Li, Z., & Li, B. (2022) · Advanced NanoBiomed Research · doi:10.1002/anbr.202200082 · Cited by 25
CRISPR applications for Duchenne muscular dystrophy: From animal models to potential therapies
Chey, Y. C. J., Arudkumar, J., Aartsma-Rus, A. M., Adikusuma, F., & Thomas, P. Q. (2022) · WIREs Mechanisms of Disease · doi:10.1002/wsbm.1580 · Cited by 24
Preclinical model for phenotypic correction of dystrophic epidermolysis bullosa by in vivo CRISPR-Cas9 delivery using adenoviral vectors
García, M., Bonafont, J., Martínez‐Palacio, J., Xu, R., Turchiano, G., Svensson, S., Thrasher, A. J., Larcher, F., Río, M. D., Hernández-Alcoceba, R., et al. (2022) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2022.09.005 · Cited by 25
Future Directions for Adrenal Insufficiency: Cellular Transplantation and Genetic Therapies
Graves, L. E., Torpy, D. J., Coates, P. T., Alexander, I. E., Bornstein, S. R., & Clarke, B. M. (2023) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/clinem/dgac751 · Cited by 23
You can't keep a bad idea down: Dark history, death, and potential rebirth of eugenics
Liscum, M., & Garcia, M. L. (2021) · The Anatomical Record · doi:10.1002/ar.24849 · Cited by 26
CRISPR/Cas9 for hepatitis B virus infection treatment
Cai, B., Chang, S., Tian, Y., & Zhen, S. (2023) · Immunity Inflammation and Disease · doi:10.1002/iid3.866 · Cited by 21
Targeted, safe, and efficient gene delivery to human hematopoietic stem and progenitor cells in vivo using the engineered AVID adenovirus vector platform
Yao, J., Atasheva, S., Wagner, N. J., Paolo, N. C. D., Stewart, P. L., & Shayakhmetov, D. M. (2023) · Molecular Therapy · doi:10.1016/j.ymthe.2023.10.023 · Cited by 23
CRISPR Technology for Ocular Angiogenesis
Chung, S. H., Sin, T.-N., Ngo, T., & Yiu, G. C. (2020) · Frontiers in Genome Editing · doi:10.3389/fgeed.2020.594984 · Cited by 21
Silencing hepatitis B virus covalently closed circular DNA: The potential of an epigenetic therapy approach
Singh, P., Kairuz, D., Arbuthnot, P. B., & Bloom, K. M. (2021) · World Journal of Gastroenterology · doi:10.3748/wjg.v27.i23.3182 · Cited by 19
In vivo genome editing at the albumin locus to treat methylmalonic acidemia
Schneller, J. L., Lee, C. M., Venturoni, L. E., Chandler, R. J., Li, A., Myung, S., Cradick, T. J., Hurley, A. E., Lagor, W. R., Bao, G., et al. (2021) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2021.11.004 · Cited by 20
Self-citation
Electroporation-Mediated Delivery of Cas9 Ribonucleoproteins Results in High Levels of Gene Editing in Primary Hepatocytes
Rathbone, T., Ates, I., Fernando, L. P., Addlestone, E., Lee, C. M., Richards, V. P., & Cottle, R. N. (2022) · The CRISPR Journal · doi:10.1089/crispr.2021.0134 · Cited by 23
Self-citation
CRISPR–Cas technologies in neurodegenerative disorders: mechanistic insights, therapeutic potential, and translational challenges
Yashooa, R. K., Nabi, A. Q., Smail, S. W., Azeez, S. S., Nooh, W. A., Mustafa, S. A., Al-Farha, A. A.-B., Capitanio, N., & Shekha, M. S. (2026) · Frontiers in Neurology · doi:10.3389/fneur.2025.1737468 · Cited by 21
Fast and Efficient Genome Editing of Human FOXP3+ Regulatory T Cells
Zeebroeck, L. V., Hornero, R. A., Côrte‐Real, B. F., Hamad, I., Meissner, T. B., & Kleinewietfeld, M. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2021.655122 · Cited by 22
In Vivo Hematopoietic Stem Cell Genome Editing: Perspectives and Limitations
Psatha, N., Paschoudi, K., Παπαδοπούλου, Α., & Yannaki, E. (2022) · Genes · doi:10.3390/genes13122222 · Cited by 24
Toward the Treatment of Inherited Diseases of the Retina Using CRISPR-Based Gene Editing
Hernández-Juárez, J., Rodríguez‐Uribe, G., & Borooah, S. (2021) · Frontiers in Medicine · doi:10.3389/fmed.2021.698521 · Cited by 19
Non-viral approaches for gene therapy and therapeutic genome editing across the blood–brain barrier
Xie, R., Wang, Y., Burger, J. C., Li, D., Zhu, M., & Gong, S. (2023) · Med-X · doi:10.1007/s44258-023-00004-0 · Cited by 22
Nanotechnology in Retinal Disease: Current Concepts and Future Directions
Mehta, N. J., & Mehta, S. N. (2023) · Journal of Ocular Pharmacology and Therapeutics · doi:10.1089/jop.2023.0083 · Cited by 18
Efficient and multiplexed somatic genome editing with Cas12a mice
Hebert, J. D., Xu, H., Tang, Y. J., Ruiz, P. A., Detrick, C. R., Wang, J., Hughes, N. W., Donosa, O., Siah, V. P., Andrejka, L., et al. (2025) · Nature Biomedical Engineering · doi:10.1038/s41551-025-01407-7 · Cited by 18
In vivo liver targeted genome editing as therapeutic approach: progresses and challenges
Simoni, C., Barbon, E., Muro, A. F., & Cantore, A. (2024) · Frontiers in Genome Editing · doi:10.3389/fgeed.2024.1458037 · Cited by 20
Genetic and functional correction of argininosuccinate lyase deficiency using CRISPR adenine base editors
Jalil, S., Keskinen, T., Juutila, J., Sartori-Maldonado, R., Euro, L., Suomalainen, A., Lapatto, R., Kuuluvainen, E., Hietakangas, V., Otonkoski, T., et al. (2024) · The American Journal of Human Genetics · doi:10.1016/j.ajhg.2024.03.004 · Cited by 18
New Directions in Pulmonary Gene Therapy
Vu, A., & McCray, P. B. (2020) · Human Gene Therapy · doi:10.1089/hum.2020.166 · Cited by 19
Therapeutic in vivo genome editing: innovations and challenges in rAAV vector-based CRISPR delivery
Gil, J.-S., Lee, S., & Koo, T. (2025) · Gene Therapy · doi:10.1038/s41434-025-00573-2 · Cited by 19
Reduction of Pre-Existing Adaptive Immune Responses Against SaCas9 in Humans Using Epitope Mapping and Identification
Shen, X., Lin, Q., Liang, Z., Wang, J., Yang, X., Liang, Y., Liang, H., Pan, H., Yang, J., Zhu, Y., et al. (2022) · The CRISPR Journal · doi:10.1089/crispr.2021.0142 · Cited by 16
Doxycycline-dependent Cas9-expressing pig resources for conditional in vivo gene nullification and activation
Jin, Q., Liu, X., Zhuang, Z., Huang, J., Gou, S., Shi, H., Zhao, Y., Ouyang, Z., Liu, Z., Li, L., et al. (2023) · Genome biology · doi:10.1186/s13059-023-02851-x · Cited by 15
Liver-directed gene-based therapies for inborn errors of metabolism
Piccolo, P., Rossi, A., & Brunetti‐Pierri, N. (2020) · Expert Opinion on Biological Therapy · doi:10.1080/14712598.2020.1817375 · Cited by 16

Authors: 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., & Lagor, W. R.
Affiliations: Rice University (US)
Baylor College of Medicine (US)
Corresponding author: Bao, Gang
Beeton, Christine
Lagor, William R.
ORCID: Li, Ang
Tanner, Mark R.
Lee, Ciaran M.
Hurley, Ayrea E.
Jarrett, Kelsey E.
Doerfler, Alexandria M.
Bao, Gang
Beeton, Christine
Lagor, William R.
Venue: Molecular Therapy
Publisher: Elsevier BV
Published: 2020-06
Volume: 28
Issue: 6
Pages: 1432-1441
ISSN: 1525-0016 (ISSN-L)
DOI: 10.1016/j.ymthe.2020.04.017
PMC: PMC7264438
OpenAlex: W3016959969
Cited by: 230
References: 40
Citations by year: 2026: 26 · 2025: 33 · 2024: 33 · 2023: 36 · 2022: 38 · 2021: 43 · 2020: 19
FWCI: 11.02
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor)
Locations: Molecular Therapy (publishedVersion, cc-by-nc-nd)
PubMed Central (publishedVersion, cc-by-nc-nd)
PubMed (publishedVersion)
Digital Commons-TMC (Texas Medical Center) (submittedVersion)
Figshare (submittedVersion, cc-by)
Health Research Alliance (cc-by)
Funders: American Heart Association (19POST34430092, 19PRE34380467, GM08231, HL007676, RR024574)
Cancer Prevention and Research Institute of Texas (RR140081, RP180672)
National Institutes of Health (RR024574, HL007676, GM08231, CA125123, UG3HL151545, HL132840, U42OD026645)
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, SaCas9, pre-existing immunity, neutralizing antibodies, T cell response, hepatocyte apoptosis, compensatory liver regeneration, SpCas9
MeSH terms: Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Dependovirus, Animals, Genetic Vectors, Humans, Immunization, Immunologic Memory, Biomarkers, Gene Expression, Immunophenotyping, T-Lymphocyte Subsets, Transgenes, Hepatocytes, Gene Order, Mice, Clustered Regularly Interspaced Short Palindromic Repeats

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

AAV-CRISPR Gene Editing Is Negated by Pre-existing Immunity to Cas9

Abstract

Overview

Integrity signals

Figures

Verification

Claims the paper makes

Claims other papers make about this one

Self-check

Citations

Cites

Cited by

Related

Metadata

Record

About these checks