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PMID: 24487629 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Sequence-specific inhibition of microRNA via CRISPR/CRISPRi system.

Scientific reports ·Vol. 4 ·2014-02-03 ·Pages 3943

Zhao Y, Dai Z, Liang Y, Yin M, Ma K, He M, Ouyang H, Teng CB

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.

MeSH Terms
3T3 Cells Animals Bacterial Proteins/metabolism CRISPR-Associated Protein 9 Cell Line Clustered Regularly Interspaced Short Palindromic Repeats/genetics Endonucleases/metabolism Gene Expression Gene Targeting/methods Green Fluorescent Proteins/genetics Mice MicroRNAs/antagonists & inhibitors,genetics RNA, Guide/genetics RNA, Small Interfering/antagonists & inhibitors,genetics Recombinant Fusion Proteins/biosynthesis,genetics Swine
Chemicals
Bacterial Proteins MicroRNAs RNA, Guide RNA, Small Interfering Recombinant Fusion Proteins enhanced green fluorescent protein Green Fluorescent Proteins CRISPR-Associated Protein 9 Cas9 protein, Francisella novicida Endonucleases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Zhao Yicheng
1] College of life science, Northeast Forestry University, Harbin, China [2] Department of Biochemistry, College of Animal Sciences, Jilin University, China [3].
Dai Zhen
1] Department of Biochemistry, College of Animal Sciences, Jilin University, China [2].
Liang Yang
1] College of life science, Northeast Forestry University, Harbin, China [2].
Yin Ming
College of Veterinary Medicine, Jilin University, China.
Ma Kuiying
College of Veterinary Medicine, Jilin University, China.
He Mei
College of life science, Northeast Forestry University, Harbin, China.
Ouyang Hongsheng
Department of Biochemistry, College of Animal Sciences, Jilin University, China.
Teng Chun-Bo
College of life science, Northeast Forestry University, Harbin, China.
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Article Info
Journal
Scientific reports
Abbr.
Sci Rep
ISSN
2045-2322
DOI
10.1038/srep03943
Published
2014-02-03
Epub
2014-00-03
Pages
3943
Language
English
Region
England
NLM ID
101563288
PMCID
PMC3909901
Subset
IM
External Links
PubMed source DOI: 10.1038/srep03943 PMC Full Text View journal details
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