高浓度原核注射后小鼠中高效的Cpf1介导的基因靶向
Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection.
作者信息
Watkins-Chow Dawn E, Varshney Gaurav K, Garrett Lisa J, Chen Zelin, Jimenez Erin A, Rivas Cecilia, Bishop Kevin S, Sood Raman, Harper Ursula L, Pavan William J, Burgess Shawn M
机构信息
Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
Functional and Chemical Genomics Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
出版信息
G3 (Bethesda). 2017 Feb 9;7(2):719-722. doi: 10.1534/g3.116.038091.
Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.
Cpf1已成为Cas9 RNA引导核酸酶的替代物。在此我们表明，在小鼠中使用Cpf1的基因靶向率能够达到甚至超过Cas9的靶向率（接近100%靶向），但需要更高浓度的mRNA和向导。我们还证明，共注射两个具有紧密靶向位点的向导可导致协同基因组切割，即便其中一个向导的切割活性极低。
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