RNA-guided DNA insertion with CRISPR-associated transposases.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 31171706.
- Also identified by DOI 10.1126/science.aax9181 and PMC identifier 6659118.
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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 <i>Scytonema hofmanni</i> (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 <i>Escherichia coli</i> 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.
Medical subject headings
- CRISPR-Cas Systems
- Cyanobacteria
- DNA Transposable Elements
- Gene Editing
- Mutagenesis, Insertional
- RNA, Guide, CRISPR-Cas Systems
- Transposases