利用RNA干扰技术沉默基因表达在本科实验教学中的设计与实践

doi:10.16288/j.yczz.18-343

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Hereditas(Beijing) ›› 2019, Vol. 41 ›› Issue (7): 653-661.doi: 10.16288/j.yczz.18-343

• Genetics Teaching • Previous Articles

Laboratory design and practice for undergraduates: Using RNAi to modulate gene expression

Yakun Song,Min Zhang,Qiaochu Wang,Yuli Peng,Fangxing Jia,Chunhong Yu()

Life Science Experimential Teaching Center, Department of Biology, Southern University of Science and Technology, Shenzhen 518055, China

Received:2019-01-18 Revised:2019-05-12 Online:2019-07-20 Published:2019-06-04
Contact: Yu Chunhong E-mail:yuch@sustech.edu.cn
Supported by:
Supported by the Teaching Reform Projects of Southern University of Science and Technology(ZLGC201603);Supported by the Teaching Reform Projects of Southern University of Science and Technology(JG201604);Supported by the Teaching Reform Projects of Southern University of Science and Technology(SJZLGC201701)

Abstract

Abstract:

RNA interference is a gene silencing phenomenon mediated by short double-stranded RNAs, which has become a widely used research technology for reverse genetics. In order to make students understand the technology better, the students were required to select target genes, to design small interfering RNAs (siRNAs) and primers, and then to test the effect of gene silencing mediated by siRNAs. Taking the fifth group in 2018 as an example, Mus musculus acyl-CoA synthetase long-chain family member 1 (Acsl1) was selected as the target gene, two pairs of siRNAs targeting Acsl1 mRNA were designed and transfected into 3T3-L1 by electroporation, then the total RNAs were extracted and synthesized to cDNA, and the expression levels of mRNAs were finally tested by relative quantitative PCR. The results showed that both pairs of siRNAs had more than 60% silencing effects. In the past three years, about 83% of the students completed all the experiments successfully and screened out at least a pair of effective siRNA. This teaching practice for undergraduates enhances students' understanding of RNA interference principle and technology, and exercises students’ lab experience and scientific research ability.

Key words: RNA interference (RNAi), siRNA design, laboratory course, undergraduate education

Yakun Song,Min Zhang,Qiaochu Wang,Yuli Peng,Fangxing Jia,Chunhong Yu. Laboratory design and practice for undergraduates: Using RNAi to modulate gene expression[J]. Hereditas(Beijing), 2019, 41(7): 653-661.

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