Rapid characterization of Cas endonuclease systems, PAM sequences and guide RNA elements
申请人:PIONEER HI-BRED INTERNATIONAL, INC.
公开号:US11371050B2
公开(公告)日:2022-06-28
Compositions and methods are provided for rapid characterization of Cas endonuclease systems and the elements comprising such systems, including, but not limiting to, rapid characterization of PAM sequences, guide RNA elements and Cas endonucleases. Type II Cas9 endonuclease systems originating from Brevibacillus laterosporus, Lactobacillus reuteri Mlc3, Lactobacillus rossiae DSM 15814, Pediococcus pentosaceus SL4, Lactobacillus nodensis JCM 14932, Sulfurospirillum sp. SCADC, Bifidobacterium thermophilum DSM 20210, Loktanella vestfoldensis, Sphingomonas sanxanigenens NX02, Epilithonimonas tenax DSM 16811, Sporocytophaga myxococcoides are described herein. The present disclosure also describes methods for genome modification of a target sequence in the genome of a cell, for gene editing, and for inserting a polynucleotide of interest into the genome of a cell.
本文提供了用于快速鉴定 Cas 内切酶系统及其组成元件的组合物和方法,包括但不限于快速鉴定 PAM 序列、引导 RNA 元件和 Cas 内切酶。II 型 Cas9 内切酶系统源自 Brevibacillus laterosporus、Lactobacillus reuteri Mlc3、Lactobacillus rossiae DSM 15814、Pediococcus pentosaceus SL4、Lactobacillus nodensis JCM 14932、Sulfurospirillum sp.在此描述的有:SCADC、嗜热双歧杆菌 DSM 20210、Loktanella vestfoldensis、Sphingomonas sanxanigenens NX02、Epilithonimonas tenax DSM 16811、Sporocytophaga myxoccoides。本公开内容还描述了对细胞基因组中的目标序列进行基因组修饰、基因编辑以及将感兴趣的多核苷酸插入细胞基因组的方法。