| RRC ID |
43892
|
| 著者 |
Takahashi M, Elbarbary RA, Abe M, Sato M, Yoshida T, Yamada Y, Tamura M, Nashimoto M.
|
| タイトル |
Elimination of specific miRNAs by naked 14-nt sgRNAs.
|
| ジャーナル |
PLoS One
|
| Abstract |
tRNase Z(L)-utilizing efficacious gene silencing (TRUE gene silencing) is a newly developed technology to suppress mammalian gene expression. TRUE gene silencing works on the basis of a unique enzymatic property of mammalian tRNase Z(L), which is that it can recognize a pre-tRNA-like or micro-pre-tRNA-like complex formed between target RNA and artificial small guide RNA (sgRNA) and can cleave any target RNA at any desired site. There are four types of sgRNA, 5'-half-tRNA, RNA heptamer, hook RNA, and ~14-nt linear RNA. Here we show that a 14-nt linear-type sgRNA against human miR-16 can guide tRNase Z(L) cleavage of miR-16 in vitro and can downregulate the miR-16 level in HEK293 cells. We also demonstrate that the 14-nt sgRNA can be efficiently taken up without any transfection reagents by living cells and can exist stably in there for at least 24 hours. The naked 14-nt sgRNA significantly reduced the miR-16 level in HEK293 and HL60 cells. Three other naked 14-nt sgRNAs against miR-142-3p, miR-206, and miR-19a/b are also shown to downregulate the respective miRNA levels in various mammalian cell lines. Our observations suggest that in general we can eliminate a specific cellular miRNA at least by ~50% by using a naked 14-nt sgRNA on the basis of TRUE gene silencing.
|
| 巻・号 |
7(6)
|
| ページ |
e38496
|
| 公開日 |
2012-1-1
|
| DOI |
10.1371/journal.pone.0038496
|
| PII |
PONE-D-11-24813
|
| PMID |
22675569
|
| PMC |
PMC3366928
|
| MeSH |
Base Sequence
Cell Line
Cell Survival
Down-Regulation / genetics
Endoribonucleases
Gene Silencing
Humans
MicroRNAs / genetics
MicroRNAs / metabolism*
Molecular Sequence Data
Nucleotides / metabolism*
RNA, Guide, Kinetoplastida / genetics
RNA, Guide, Kinetoplastida / metabolism*
|
| IF |
2.74
|
| 引用数 |
8
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
293(RCB1637)
HeLa(RCB0007)
Jurkat(RCB0806)
HL60(RCB0041)
DAUDI |
