RNA-guided DNA insertion with CRISPR-associated transposases
Name
nihms-1037885.pdf
Description
Accepted version
Size
1.46 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Strecker, Jonathan
Ladha, Alim
Gardner, Zachary
Schmid-Burgk, Jonathan L.
Makarova, Kira S.
Koonin, Eugene V.
Zhang, Feng
Date Issued
July 2019
Journal
Science
Publisher
American Association for the Advancement of Science (AAAS)
Citation
Strecker, Jonathan et al. "RNA-guided DNA insertion with CRISPR-associated transposases." Science 365, 6448 (July 2019): 48-53 © 2019 The Authors
Version
Author's final manuscript
Abstract
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.
MIT Department
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution-Noncommercial-Share Alike
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1126/science.aax9181
