Reprogramming an RNA-guided archaeal TnpB endonuclease for genome editing
Ying Xu, Tao Liu, Jing Wang, Binyang Xiong, Ling Liu, Nan Peng
Abstract
CRISPR-Cas systems are prokaryotic immune systems, protecting prokaryotes against invasive viruses and plasmids 1 . The CRISPR ribonucleoproteins (RNPs) recognize DNA or RNA target through a mature CRISPR RNA and cleave the target in a sequence-specific manner 2 . The discovery of the CRISPR-Cas system has promoted the development of new genome editing tools based on Cas9 3 and Cas12a 4 . Recently, new programmable endonucleases have been characterized to expand the genome engineering toolbox. The ancestors of Cas9 and Cas12 family proteins, IscB and TnpB, respectively, have been identified from the widespread IS200/IS605 and IS607 transposon families 5 , 6 , 7 . Both IscB and TnpB use a single noncoding RNA for RNA-guided cleavage of double-stranded (ds) DNA 8 , 9 , exhibiting genome editing activity in human cells 9 , 10 .