Elimination of specific miRNAs by naked 14-nt sgRNAs

Masayuki Takahashi, Reyad Elbarbary, Mayumi Abe, Mari Sato, Tetsuo Yoshida, Yoji Yamada, Masato Tamura, Masayuki Nashimoto

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish (US)
Article numbere38496
JournalPloS one
Volume7
Issue number6
DOIs
StatePublished - Jun 4 2012

Fingerprint

Guide RNA
MicroRNAs
RNA
Gene Silencing
Genes
HEK293 Cells
RNA Precursors
gene silencing
Down-Regulation
Cells
Hooks
HL-60 Cells
microRNA
Transfer RNA
Gene expression
Transfection
Technology
Gene Expression
Cell Line
cells

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Takahashi, M., Elbarbary, R., Abe, M., Sato, M., Yoshida, T., Yamada, Y., ... Nashimoto, M. (2012). Elimination of specific miRNAs by naked 14-nt sgRNAs. PloS one, 7(6), [e38496]. https://doi.org/10.1371/journal.pone.0038496
Takahashi, Masayuki ; Elbarbary, Reyad ; Abe, Mayumi ; Sato, Mari ; Yoshida, Tetsuo ; Yamada, Yoji ; Tamura, Masato ; Nashimoto, Masayuki. / Elimination of specific miRNAs by naked 14-nt sgRNAs. In: PloS one. 2012 ; Vol. 7, No. 6.
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Takahashi, M, Elbarbary, R, Abe, M, Sato, M, Yoshida, T, Yamada, Y, Tamura, M & Nashimoto, M 2012, 'Elimination of specific miRNAs by naked 14-nt sgRNAs', PloS one, vol. 7, no. 6, e38496. https://doi.org/10.1371/journal.pone.0038496

Elimination of specific miRNAs by naked 14-nt sgRNAs. / Takahashi, Masayuki; Elbarbary, Reyad; Abe, Mayumi; Sato, Mari; Yoshida, Tetsuo; Yamada, Yoji; Tamura, Masato; Nashimoto, Masayuki.

In: PloS one, Vol. 7, No. 6, e38496, 04.06.2012.

Research output: Contribution to journalArticle

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AU - Takahashi, Masayuki

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Takahashi M, Elbarbary R, Abe M, Sato M, Yoshida T, Yamada Y et al. Elimination of specific miRNAs by naked 14-nt sgRNAs. PloS one. 2012 Jun 4;7(6). e38496. https://doi.org/10.1371/journal.pone.0038496