Prediction and validation of hematopoietic stem and progenitor cell off-target editing in transplanted rhesus macaques
DC Field | Value | Language |
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dc.contributor.author | AlJanahi Aisha A. | - |
dc.contributor.author | Lazzarotto Cicera R. | - |
dc.contributor.author | Chen Shirley | - |
dc.contributor.author | Shin Tae-Hoon | - |
dc.contributor.author | Cordes Stefan | - |
dc.contributor.author | Fan Xing | - |
dc.contributor.author | Jabara Isabel | - |
dc.contributor.author | Zhou Yifan | - |
dc.contributor.author | Young David J. | - |
dc.contributor.author | Lee, Byung Chul | - |
dc.contributor.author | Yu Kyung-Rok | - |
dc.contributor.author | Li Yuesheng | - |
dc.contributor.author | Toms Bradley | - |
dc.contributor.author | Tunc Ilker | - |
dc.contributor.author | Hong So Gun | - |
dc.contributor.author | Truitt Lauren L. | - |
dc.contributor.author | Klermund Julia | - |
dc.contributor.author | Andrieux Geoffroy | - |
dc.contributor.author | Kim Miriam Y. | - |
dc.contributor.author | Cathomen Toni | - |
dc.contributor.author | Gill Saar | - |
dc.contributor.author | Tsai Shengdar Q. | - |
dc.contributor.author | Dunbar Cynthia E. | - |
dc.date.accessioned | 2023-09-22T06:40:13Z | - |
dc.date.available | 2023-09-22T06:40:13Z | - |
dc.date.issued | 2022-01 | - |
dc.identifier.issn | 1525-0016 | - |
dc.identifier.issn | 1525-0024 | - |
dc.identifier.uri | https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/151536 | - |
dc.description.abstract | The programmable nuclease technology CRISPR-Cas9 has revolutionized gene editing in the last decade. Due to the risk of off-target editing, accurate and sensitive methods for off-target characterization are crucial prior to applying CRISPR-Cas9 therapeutically. Here, we utilized a rhesus macaque model to compare the predictive values of CIRCLE-seq, an in vitro off-target prediction method, with in silico prediction (ISP) based solely on genomic sequence comparisons. We use AmpliSeq HD error-corrected sequencing to validate offtarget sites predicted by CIRCLE-seq and ISP for a CD33 guide RNA (gRNA) with thousands of off-target sites predicted by ISP and CIRCLE-seq. We found poor correlation between the sites predicted by the two methods. When almost 500 sites predicted by each method were analyzed by error-corrected sequencing of hematopoietic cells following transplantation, 19 off-target sites revealed insertion or deletion mutations. Of these sites, 8 were predicted by both methods, 8 by CIRCLE-seq only, and 3 by ISP only. The levels of cells with these off-target edits exhibited no expansion or abnormal behavior in vivo in animals followed for up to 2 years. In addition, we utilized an unbiased method termed CAST-seq to search for translocations between the on-target site and off-target sites present in animals following transplantation, detecting one specific translocation that persisted in blood cells for at least 1 year following transplantation. In conclusion, neither CIRCLE-seq or ISP predicted all sites, and a combination of careful gRNA design, followed by screening for predicted off-target sites in target cells by multiple methods, may be required for optimizing safety of clinical development. | - |
dc.format.extent | 14 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | Nature Publishing Group | - |
dc.title | Prediction and validation of hematopoietic stem and progenitor cell off-target editing in transplanted rhesus macaques | - |
dc.type | Article | - |
dc.publisher.location | 미국 | - |
dc.identifier.doi | 10.1016/j.ymthe.2021.06.016 | - |
dc.identifier.scopusid | 2-s2.0-8511466776 | - |
dc.identifier.wosid | 000744579600003 | - |
dc.identifier.bibliographicCitation | Molecular Therapy, v.30, no.1, pp 209 - 222 | - |
dc.citation.title | Molecular Therapy | - |
dc.citation.volume | 30 | - |
dc.citation.number | 1 | - |
dc.citation.startPage | 209 | - |
dc.citation.endPage | 222 | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalResearchArea | Genetics & Heredity | - |
dc.relation.journalResearchArea | Research & Experimental Medicine | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Genetics & Heredity | - |
dc.relation.journalWebOfScienceCategory | Research & Experimental Medicine | - |
dc.subject.keywordAuthor | Ca9 | - |
dc.subject.keywordAuthor | CRISPR | - |
dc.subject.keywordAuthor | error-corrected sequencing | - |
dc.subject.keywordAuthor | gene editing | - |
dc.subject.keywordAuthor | gene therapy | - |
dc.subject.keywordAuthor | Macaque | - |
dc.subject.keywordAuthor | off-target | - |
dc.subject.keywordAuthor | translocation | - |
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