NDLI: Drug-inducible synergistic gene silencing with multiple small hairpin RNA molecules for gene function study in animal model

Content Provider	Springer Nature Link
Author	Ying, Ming Chen, Guangfeng Qiu, Yu Shi, Xiujuan Zhang, Chen Wang, Qiuke Yang, Shuzhang Lu, Lixia Yuan, Qionglan Xu, Guotong Jin, Zibing Wu, Qiang Liu, Xiaoqing
Copyright Year	2014
Abstract	Gene targeting is a critical tool for construction of disease models. However, the application of traditional homologous recombination-mediated gene knockout technology is limited by the absence of rapid frequency-guaranteed targeting methods. Although conventional small hairpin RNA (shRNA)-mediated gene silencing offers an alternative for gene targeting, its application is frequently compromised by lower expression efficiency via RNA interference compared to gene knockout. Here we provide an efficient gene targeting strategy involving drug-inducible synergistic silencing with multiple shRNA molecules. On induction, the levels of the target proteins decreased to undetectable levels in all the tested stable transgenic mammalian cell lines, including HEK293 and embryonic stem cell-derived progenies carrying shRNA silencing cassettes. In a transgenic mouse model carrying a silencing cassette targeting the rhodopsin gene, short-time inducer treatment was sufficient to ablate the rhodopsin protein in the retina, resulting in similar retinal phenotypic changes as those observed in rhodopsin mutant mice. Therefore, on a broad basis, this inducible shRNA gene targeting strategy offers a true gene knockout alternative comparable to conventional RNA interference approaches.
Starting Page	309
Ending Page	317
Page Count	9
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	24
Issue Number	2
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2014-10-01
Publisher Place	Cham
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	RNAi shRNA Gene targeting Gene silencing Drug-inducible Molecular Medicine Plant Genetics & Genomics Animal Genetics and Genomics Plant Sciences Transgenics
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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