MiCas9 increases large size gene knock-in rates and reduces undesirable on-target and off-target indel edits.
dc.contributor.author | Ma, L | |
dc.contributor.author | Ruan, J | |
dc.contributor.author | Song, J | |
dc.contributor.author | Wen, L | |
dc.contributor.author | Yang, D | |
dc.contributor.author | Zhao, J | |
dc.contributor.author | Xia, X | |
dc.contributor.author | Chen, YE | |
dc.contributor.author | Zhang, J | |
dc.contributor.author | Xu, J | |
dc.coverage.spatial | England | |
dc.date.accessioned | 2022-10-05T14:51:35Z | |
dc.date.available | 2022-10-05T14:51:35Z | |
dc.date.issued | 2020-01-01 | |
dc.identifier.issn | 2041-1723 | |
dc.identifier.issn | 2041-1723 | |
dc.identifier.uri | https://www.ncbi.nlm.nih.gov/pubmed/33247137 | |
dc.identifier.uri | https://hdl.handle.net/2027.42/174895 | en |
dc.description.abstract | Gene editing nuclease represented by Cas9 efficiently generates DNA double strand breaks at the target locus, followed by repair through either the error-prone non-homologous end joining or the homology directed repair pathways. To improve Cas9’s homology directed repair capacity, here we report the development of miCas9 by fusing a minimal motif consisting of thirty-six amino acids to spCas9. MiCas9 binds RAD51 through this fusion motif and enriches RAD51 at the target locus. In comparison to spCas9, miCas9 enhances double-stranded DNA mediated large size gene knock-in rates, systematically reduces off-target insertion and deletion events, maintains or increases single-stranded oligodeoxynucleotides mediated precise gene editing rates, and effectively reduces on-target insertion and deletion rates in knock-in applications. Furthermore, we demonstrate that this fusion motif can work as a “plug and play” module, compatible and synergistic with other Cas9 variants. MiCas9 and the minimal fusion motif may find broad applications in gene editing research and therapeutics. | |
dc.format.medium | Electronic | |
dc.language | eng | |
dc.publisher | Springer Nature | |
dc.relation.haspart | ARTN 6082 | |
dc.rights | Licence for published version: Creative Commons Attribution 4.0 International | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.subject | Amino Acid Sequence | |
dc.subject | CRISPR-Associated Protein 9 | |
dc.subject | Cell Line | |
dc.subject | Gene Editing | |
dc.subject | Gene Knock-In Techniques | |
dc.subject | Green Fluorescent Proteins | |
dc.subject | Humans | |
dc.subject | INDEL Mutation | |
dc.subject | Rad51 Recombinase | |
dc.subject | Vascular Endothelial Growth Factor A | |
dc.title | MiCas9 increases large size gene knock-in rates and reduces undesirable on-target and off-target indel edits. | |
dc.type | Article | |
dc.identifier.pmid | 33247137 | |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/174895/2/MiCas9 increases large size gene knock-in rates and reduces undesirable on-target and off-target indel edits.pdf | |
dc.identifier.doi | 10.1038/s41467-020-19842-2 | |
dc.identifier.doi | https://dx.doi.org/10.7302/6524 | |
dc.identifier.source | Nat Commun | |
dc.description.version | Published version | |
dc.date.updated | 2022-10-05T14:51:26Z | |
dc.identifier.orcid | 0000-0003-2449-8651 | |
dc.identifier.orcid | 0000-0003-2357-7825 | |
dc.identifier.volume | 11: 6082. | |
dc.identifier.issue | 1 | |
dc.identifier.startpage | 6082 | |
dc.identifier.name-orcid | Ma, L | |
dc.identifier.name-orcid | Ruan, J | |
dc.identifier.name-orcid | Song, J | |
dc.identifier.name-orcid | Wen, L | |
dc.identifier.name-orcid | Yang, D; 0000-0003-2449-8651 | |
dc.identifier.name-orcid | Zhao, J | |
dc.identifier.name-orcid | Xia, X | |
dc.identifier.name-orcid | Chen, YE; 0000-0003-2357-7825 | |
dc.identifier.name-orcid | Zhang, J | |
dc.identifier.name-orcid | Xu, J | |
dc.working.doi | 10.7302/6524 | en |
dc.owningcollname | Internal Medicine, Department of |
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