Abstract
CRISPR advances genome engineering by directing endonuclease sequence specificity with a guide RNA molecule (gRNA). For precisely targeting a gene for modification, each genetic construct requires a unique gRNA. By generating a gRNA against the flippase recognition target (FRT) site, a common genetic element shared by multiple genetic collections, CRISPR-FRT circumvents this design constraint to provide a broad platform for fast, scarless, off-the-shelf genome engineering.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Binding Sites / genetics
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CRISPR-Cas Systems*
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DNA Nucleotidyltransferases / genetics
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DNA Nucleotidyltransferases / metabolism*
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DNA, Bacterial / genetics
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DNA, Bacterial / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Editing / methods*
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Gene Knockout Techniques
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Genome, Bacterial / genetics
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Models, Genetic
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Mutation
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RNA, Guide / genetics
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RNA, Guide / metabolism*
Substances
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DNA, Bacterial
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RNA, Guide
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DNA Nucleotidyltransferases
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Site-specific recombinase