摘要
Dear editor.The remarkably diversified CRISPR-Cas systems in nature have provided unlimited valuable resources to develop genome editing tools that are revolutionizing the fields of biotechnology and medicine.However,due to their microbial origin,the activity of most naturally occurring CRISPR-Cas nucleases is relatively poor in mammalian cells(Ran et al.,2015).Thus,improving the mammalian genome editing efficiency becomes the priority for harnessing more CRISPRCas systems for widespread applications.Rational protein engineering serves as a powerful approach to enhance the catalytic activity of enzymes.Whereas,this approach largely relies on proteinic three-dimension(3D)structure information as guide for design.The fact is that of the large numbers of the CRISPR-Cas systems discovered in recent years,only a small number of them with the 3D structures were reported.To bypass this limitation,here,we report an efficient and simple strategy for rational engineering of Cas nucleases without the need of 3D structure information and successfully optimized nucleases from Cas9 and Cas12 families.
基金
supported by the National Key Research and Development Program(2020YFA0707900,2018YFE0201100,2019YFA0110800 to W.L.,2018YFA0108400,2019YFA0903800 to Q.Z.)
the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16030403 to W.L.)
the National Natural Science Foundation of China(31621004 to Q.Z.and W.L.)
the CAS Project for Young Scientists in Basic Research(YSBR-012 to W.L.)。
