RNA-guided DNA insertion with CRISPR-associated transposases
Author(s)Strecker, Jonathan; Ladha, Alim; Gardner, Zachary; Schmid-Burgk, Jonathan L.; Makarova, Kira S.; Koonin, Eugene V.; Zhang, Feng; ... Show more Show less
MetadataShow full item record
CRISPR-Cas nucleases are powerful tools for manipulating nucleic acids; however, targeted insertion of DNA remains a challenge, as it requires host cell repair machinery. Here we characterize a CRISPR-associated transposase from cyanobacteria Scytonema hofmanni (ShCAST) that consists of Tn7-like transposase subunits and the type V-K CRISPR effector (Cas12k). ShCAST catalyzes RNA-guided DNA transposition by unidirectionally inserting segments of DNA 60 to 66 base pairs downstream of the protospacer. ShCAST integrates DNA into targeted sites in the Escherichia coli genome with frequencies of up to 80% without positive selection. This work expands our understanding of the functional diversity of CRISPR-Cas systems and establishes a paradigm for precision DNA insertion.
DepartmentMcGovern Institute for Brain Research at MIT; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; Massachusetts Institute of Technology. Department of Biological Engineering
American Association for the Advancement of Science (AAAS)
Strecker, Jonathan et al. "RNA-guided DNA insertion with CRISPR-associated transposases." Science 365, 6448 (July 2019): 48-53 © 2019 The Authors
Author's final manuscript