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Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Sojung | - |
dc.contributor.author | Kim, Daesik | - |
dc.contributor.author | Cho, Seung Woo | - |
dc.contributor.author | Kim, Jungeun | - |
dc.contributor.author | Kim, Jin-Soo | - |
dc.date.accessioned | 2020-04-27T12:39:05Z | - |
dc.date.available | 2020-04-27T12:39:05Z | - |
dc.date.created | 2020-03-23 | - |
dc.date.created | 2020-03-23 | - |
dc.date.issued | 2014-06 | - |
dc.identifier.citation | Genome Research, Vol.24 No.6, pp.1012-1019 | - |
dc.identifier.issn | 1088-9051 | - |
dc.identifier.other | 93168 | - |
dc.identifier.uri | https://hdl.handle.net/10371/165637 | - |
dc.description.abstract | RNA-guided engineered nucleases (RGENs) derived from the prokaryotic adaptive immune system known as CRISPR clustered, regularly interspaced, short palindromic repeat)/Cas (CRISPR-associated) enable genome editing in human cell lines, animals, and plants, but are limited by off-target effects and unwanted integration of DNA segments derived from plasmids encoding Cas9 and guide RNA at both on-target and off-target sites in the genome. Here, we deliver purified recombinant Cas9 protein and guide RNA into cultured human cells including hard-to-transfect fibroblasts and pluripotent stem cells. RGEN ribonucleoproteins (RNPs) induce site-specific mutations at frequencies of up to 79%, while reducing off-target mutations associated with plasmid transfection at off-target sites that differ by one or two nucleotides from on-target sites. RGEN RNPs cleave chromosomal DNA almost immediately after delivery and are degraded rapidly in cells, reducing off-target effects. Furthermore, RNP delivery is less stressful to human embryonic stem cells, producing at least twofold more colonies than does plasmid transfection. | - |
dc.language | 영어 | - |
dc.publisher | Cold Spring Harbor Laboratory Press | - |
dc.title | Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins | - |
dc.type | Article | - |
dc.contributor.AlternativeAuthor | 김진수 | - |
dc.identifier.doi | 10.1101/gr.171322.113 | - |
dc.citation.journaltitle | Genome Research | - |
dc.identifier.wosid | 000336662200012 | - |
dc.identifier.scopusid | 2-s2.0-84901834420 | - |
dc.citation.endpage | 1019 | - |
dc.citation.number | 6 | - |
dc.citation.startpage | 1012 | - |
dc.citation.volume | 24 | - |
dc.identifier.sci | 000336662200012 | - |
dc.description.isOpenAccess | Y | - |
dc.contributor.affiliatedAuthor | Kim, Jin-Soo | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.subject.keywordPlus | ZINC-FINGER NUCLEASES | - |
dc.subject.keywordPlus | GENE KNOCKOUT | - |
dc.subject.keywordPlus | HOMOLOGOUS RECOMBINATION | - |
dc.subject.keywordPlus | CAENORHABDITIS-ELEGANS | - |
dc.subject.keywordPlus | DNA | - |
dc.subject.keywordPlus | SPECIFICITY | - |
dc.subject.keywordPlus | SYSTEMS | - |
dc.subject.keywordPlus | CRISPR/CAS | - |
dc.subject.keywordPlus | MICE | - |
dc.subject.keywordPlus | ENDONUCLEASES | - |
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