Abstract
Here, we report a convenient and efficient miRNA inhibition strategy employing the CRISPR system. Using specifically designed gRNAs, miRNA gene has been cut at a single site by Cas9, resulting in knockdown of the miRNA in murine cells. Using a modified CRISPR interference system (CRISPRi), inactive Cas9 can reversibly prevent the expression of both monocistronic miRNAs and polycistronic miRNA clusters. Furthermore, CRISPR/CRISPRi is also capable of suppressing genes in porcine cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Animals
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Bacterial Proteins / metabolism
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CRISPR-Associated Protein 9
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Cell Line
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Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
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Endonucleases / metabolism
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Gene Expression
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Gene Targeting / methods*
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Green Fluorescent Proteins / genetics
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Mice
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MicroRNAs / antagonists & inhibitors*
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MicroRNAs / genetics
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RNA, Guide, CRISPR-Cas Systems / genetics
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RNA, Small Interfering / antagonists & inhibitors*
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RNA, Small Interfering / genetics
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Swine
Substances
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Bacterial Proteins
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MicroRNAs
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RNA, Guide, CRISPR-Cas Systems
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RNA, Small Interfering
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Recombinant Fusion Proteins
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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CRISPR-Associated Protein 9
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Cas9 protein, Francisella novicida
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Endonucleases