Rescue of high-specificity Cas9 variants using sgRNAs with matched 5 ' nucleotides

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Title
Rescue of high-specificity Cas9 variants using sgRNAs with matched 5 ' nucleotides
Author(s)
Sojung Kim; Taegeun Bae; Jaewoong Hwang; Jin-Soo Kim
Publication Date
2017-11
Journal
GENOME BIOLOGY, v.18, no.1, pp.218 -
Publisher
BIOMED CENTRAL LTD
Abstract
We report that engineered Cas9 variants with improved specificity-eCas9-1.1 and Cas9-HF1-are often poorly active in human cells, when complexed with single guide RNAs (sgRNAs) with a mismatch at the 5' terminus, relative to target DNA sequences. Because the nucleotide at the 5' end of sgRNAs, expressed under the control of the commonly-used U6 promoter, is fixed to a guanine, these attenuated Cas9 variants are not useful at many target sites. By using sgRNAs with matched 5' nucleotides, produced by linking them to a self-cleaving ribozyme, the editing activity of Cas9 variants can be rescued without sacrificing high specificity. © The Author(s). 2017
URI
https://pr.ibs.re.kr/handle/8788114/4120
ISSN
1474-760x
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
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