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Weng Y, Shi Y, Xia X, et al. A multi-shRNA vector enhances the silencing efficiency of exogenous and endogenous genes in human cells. Oncol Lett. 2017;13(3):1553-1562doi: 10.3892/ol.2017.5672.
Weng, Y., Shi, Y., Xia, X., Zhou, W., Wang, H., & Wang, C. (2017). A multi-shRNA vector enhances the silencing efficiency of exogenous and endogenous genes in human cells. Oncology letters, 13(3), 1553-1562. https://doi.org/10.3892/ol.2017.5672
Weng, Yanjie, et al. "A multi-shRNA vector enhances the silencing efficiency of exogenous and endogenous genes in human cells." Oncology letters vol. 13,3 (2017): 1553-1562. doi: https://doi.org/10.3892/ol.2017.5672
Weng Y, Shi Y, Xia X, Zhou W, Wang H, Wang C. A multi-shRNA vector enhances the silencing efficiency of exogenous and endogenous genes in human cells. Oncol Lett. 2017 Mar;13(3):1553-1562. doi: 10.3892/ol.2017.5672. Epub 2017 Feb 02. PMID: 28454290; PMCID: PMC5403481.
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Oncol Lett. 2017 Mar;13(3):1553-1562. doi: 10.3892/ol.2017.5672. Epub 2017 Feb 02.

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

Oncology letters

Yanjie Weng, Ying Shi, Xi Xia, Wenjuan Zhou, Hongyan Wang, Changyu Wang

Affiliations

  1. Cancer Biology Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.
  2. Department of Gynecology and Obstetrics, Affiliated Shenzhen Nanshan Hospital, Guangdong Medical College, Shenzhen, Guangdong 518052, P.R. China.

PMID: 28454290 PMCID: PMC5403481 DOI: 10.3892/ol.2017.5672

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.

Keywords: Akt2; DsRed; multi-site short hairpin RNA vector

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