Abstract
We demonstrate that by altering the length of Cas9-associated guide RNA (gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein.
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
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CRISPR-Associated Proteins / genetics
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CRISPR-Cas Systems / genetics*
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Flow Cytometry
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Fluorescent Dyes / analysis
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Gene Deletion
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Genes, Reporter
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Genetic Engineering / methods
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Genetic Vectors
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Genome
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HEK293 Cells
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Humans
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Microscopy, Fluorescence
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Mutagenesis
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Mutation
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RNA Editing
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RNA, Guide, CRISPR-Cas Systems
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Transcription, Genetic
Substances
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CRISPR-Associated Proteins
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Fluorescent Dyes