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 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.
1] College of life science, Northeast Forestry University, Harbin, China [2] Department of Biochemistry, College of Animal Sciences, Jilin University, China [3].
Yicheng
Y
Zhao
1] Department of Biochemistry, College of Animal Sciences, Jilin University, China [2].
Zhen
Z
Dai
1] College of life science, Northeast Forestry University, Harbin, China [2].
Yang
Y
Liang
College of Veterinary Medicine, Jilin University, China.
Ming
M
Yin
College of Veterinary Medicine, Jilin University, China.
Kuiying
K
Ma
College of life science, Northeast Forestry University, Harbin, China.
Mei
M
He
Department of Biochemistry, College of Animal Sciences, Jilin University, China.
Hongsheng
H
Ouyang
College of life science, Northeast Forestry University, Harbin, China.
Chun-Bo
CB
Teng
D016475:3T3 Cells; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D000076987:CRISPR-Associated Protein 9; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004720:Endonucleases / Q000378:metabolism; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000235:genetics; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D034741:RNA, Small Interfering / Q000037:antagonists & inhibitors* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013552:Swine
D001426:Bacterial Proteins; D035683:MicroRNAs; D000094704:RNA, Guide, CRISPR-Cas Systems; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004720:Endonucleases