Sequence-specific inhibition of microRNA via CRISPR/CRISPRi system

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...

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Veröffentlicht in:	Scientific reports 2014-02, Vol.4 (1), p.3943-3943, Article 3943
Hauptverfasser:	Zhao, Yicheng, Dai, Zhen, Liang, Yang, Yin, Ming, Ma, Kuiying, He, Mei, Ouyang, Hongsheng, Teng, Chun-Bo
Format:	Artikel
Sprache:	eng
Schlagworte:	13/100 13/109 13/31 13/44 13/89 38/22 38/23 3T3 Cells 42/41 45/90 631/337/505 631/61/185 Animals Bacterial Proteins - metabolism Cell Line Clustered Regularly Interspaced Short Palindromic Repeats - genetics CRISPR CRISPR-Associated Protein 9 Endonucleases - metabolism Flow cytometry Gene Expression Gene Targeting - methods Genomes Green Fluorescent Proteins - genetics Humanities and Social Sciences Mice MicroRNAs MicroRNAs - antagonists & inhibitors MicroRNAs - genetics miRNA multidisciplinary Mutation Proteins Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - genetics RNA polymerase RNA, Guide, CRISPR-Cas Systems - genetics RNA, Small Interfering - antagonists & inhibitors RNA, Small Interfering - genetics Science Swine
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Zusammenfassung:	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.
ISSN:	2045-2322 2045-2322
DOI:	10.1038/srep03943