Rajendran Arivazhagan, Shigi Narumi, Sumaoka Jun, Komiyama Makoto
Institute of Advanced Energy, Kyoto University, Kyoto, Japan.
Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, Ibaraki, Japan.
Curr Protoc Nucleic Acid Chem. 2019 Mar;76(1):e72. doi: 10.1002/cpnc.72. Epub 2019 Feb 5.
By combining a pair of pseudo-complementary peptide nucleic acids (pcPNAs) with S1 nuclease, a novel tool to cut DNA at a predetermined site can be obtained. Complementary pcPNAs invade the DNA duplex and base pair to each strand of a target site, creating single-stranded regions that are cleaved by S1 nuclease. The scission site can be freely modulated by the design of pcPNAs. This method can be used to cleave a single site in the human genome. This protocol presents experimental details for site-selective scission using this versatile new tool. © 2019 by John Wiley & Sons, Inc.
通过将一对假互补肽核酸(pcPNA)与S1核酸酶相结合,可以获得一种在预定位点切割DNA的新型工具。互补的pcPNA侵入DNA双链体并与靶位点的每条链碱基配对,形成被S1核酸酶切割的单链区域。切割位点可通过pcPNA的设计自由调节。该方法可用于切割人类基因组中的单个位点。本方案介绍了使用这种多功能新工具进行位点选择性切割的实验细节。© 2019约翰威立国际出版公司