Chicken 7SK promoter drives efficient shRNA transcription with species specificity.
To extend the use of RNAi in chicken, we have developed a RNA interference (RNAi) system using a shortened chicken 7SK (ch7SK) promoter. The results stated that the cloned ch7SK promoter includes multiple Oct-1 motifs, SPH domain, PSE and TATA box, without CACCC box. All RNAi groups driven by ch7SK promoter showed significant mean fluorescence intensity (MFI) reduction. In the pch7SK-shEGFP transfected DF-EGFP cell culture, the MFI reduction ratio was smaller than the pmU6-shEGFP did. In the pmU6-shEGFP transfected Vero-EGFP cell culture, the MFI was reduced significantly than the pch7SK-shEGFP did. In summary, the essential part of ch7SK promoter was capable of efficiently expressing shRNAs with relatively different interfering degrees in avian and mammalian cells, respectively. Our results suggest that ch7SK promoter is an efficient alternative to commercially mouse U6 promoter in shRNA expression with chicken cells, and provide references for furthering functional genome analysis and disease resistant breeding in chicken.
Key Laboratory of Animal Biotechnology of National Ministry of Agriculture, Institute of Veterinary Immunology,and Research Laboratory of Virology, Immunology & Bioinformatics, Division of Veterinary Microbiology & Virology, Department of Preventative Medicine, College of Veterinary Medicine,and Investigation Group of Molecular Virology, Immunology, Oncology & System Biology, Center for Bioinformatics, Northwest A&F University, Yangling 712100, Shaanxi, China; Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, State Key Laboratory of Veterinary Biotechnology, Harbin, Heilongjiang, China.
Xinyu
X
Chen
Xiaomin
X
Liu
Wei
W
Wang
Jianchao
J
Wang
Qingwen
Q
Meng
Deli
D
Zhang
D000818:Animals; D002645:Chickens / Q000235:genetics; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D055785:Gene Knockdown Techniques / Q000662:veterinary; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics* / Q000502:physiology; D013045:Species Specificity; D014158:Transcription, Genetic / Q000235:genetics* / Q000502:physiology; D014709:Vero Cells
D034741:RNA, Small Interfering
Ch7SK promoter; Lentivirus; MFI; RNA interference