Expansion of CRISPR/Cas9 genome targeting sites in zebrafish by Csy4-based RNA processing
Dear Editor,With its advantages of simple design and cost-efficiency,the CRISPR/Cas9 technology has been widely adapted for genome editing in different species including zebrafish ”1”.In zebrafish studies,guide RNA (gRNA) is usually produced via in vitro transcription followed by microinjection with Cas9 mRNA into embryos.The vectors currently used for production of gRNA contain either a T7 or SP6 promoter in vitro or U6 promoter in vivo.Among these,T7 promoter is most popularly used due to its high efficiency and,therefore,limits the gRNA targeting sites to an optimal ”GG-N18-NGG” format ”2”.
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This work was in part supported by funding from Science and Technology Program of Shenzhen,China CXZZ20140903101756864.
2015-10-23(万方平台首次上网日期,不代表论文的发表时间)
1074-1077