Rescue of high-specificity Cas9 variants using sgRNAs with matched 5’ nucleotides
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...
Paper Details
Title
Rescue of high-specificity Cas9 variants using sgRNAs with matched 5’ nucleotides
Published Date
Nov 15, 2017
Journal
Volume
18
Issue
1
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