A multi-shRNA vector enhances the silencing efficiency of exogenous and endogenous genes in human cells.

Abstract:

:RNA interference (RNAi) is a powerful technology for suppressing gene function. In most studies, small interfering RNAs (siRNAs) consist of one short hairpin RNA (shRNA) and, therefore, are often unable to achieve loss-of-function of their target genes. In the current study, an RNAi vector containing three shRNAs under the control of three RNA polymerase III U6 promoters was constructed. RNAi vectors containing one or two shRNAs were generated for comparisons. A pilot study targeting exogenously expressed DsRed in the HEK293 cell line revealed promising effects and a high selectivity for the multi-shRNA RNAi vector. Akt2 is constitutively expressed in cultured SKOV3 human ovarian cancer cells, and the multi-shRNA RNAi vector showed a strong efficiency for downregulating the expression of Akt2 in these cells, with no apparent interferon response. In addition, the Akt2-3shRNA vector, containing three shRNAs targeting Akt2, showed the best effect of all the shRNA vectors in reversing paclitaxel-induced resistance in SKOV3 cells. This study developed a widely applicable resource for enhancing the efficiency of gene silencing and a novel technique for performing complex loss-of-function screens in mammalian cells.

journal_name

Oncol Lett

journal_title

Oncology letters

authors

Weng Y,Shi Y,Xia X,Zhou W,Wang H,Wang C

doi

10.3892/ol.2017.5672

subject

Has Abstract

pub_date

2017-03-01 00:00:00

pages

1553-1562

issue

3

eissn

1792-1074

issn

1792-1082

pii

OL-0-0-5672

journal_volume

13

pub_type

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