【摘 要】
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Dear editorThe clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated proteins 9 (Cas9) systems are powerful tools for gen
【机 构】
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State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of
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Dear editorThe clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated proteins 9 (Cas9) systems are powerful tools for gene editing.Ribonucleoprotein (RNP) complex composed of Cas9 protein and sgRNA binds to specific genomic loci and generate DNA double strand breaks.While plasmids expressing Cas9 protein and sgRNA are routinely transfected into various cell lines to perform gene editing (Cong et al.,2013;Mali et al.,2013;Ran et al.,2013),direct delivery of Cas9-sgRNA RNP has shown higher efficiency and lower off-target effects (Kim et al.,2014),especially in human primary cells such as T cells (Hendel et al.,2015;Schumann et al.,2015).SgRNA can be generated by either in vitro transcription (IVT) or chemical synthesis.IVT is widely used to generate sgRNAs,since it can be easily performed in most labs.
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