RNA-guided DNA insertion with CRISPR-associated transposases

RNA-guided DNA insertion with CRISPR-associated transposases

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 tra...

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Bibliographic Details
Main Authors: Strecker, Jonathan, Ladha, Alim, Gardner, Zachary, Schmid-Burgk, Jonathan L., Makarova, Kira S., Koonin, Eugene V., Zhang, Feng
Other Authors: McGovern Institute for Brain Research at MIT
Format: Article
Language:English
Published: American Association for the Advancement of Science (AAAS) 2020
Online Access:https://hdl.handle.net/1721.1/125207
Description
Summary: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.

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