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3型聚合酶III启动子+1位置周围核苷酸的突变：对转录活性和起始位点使用的影响。

Mutation of nucleotides around the +1 position of type 3 polymerase III promoters: The effect on transcriptional activity and start site usage.

作者信息

Gao Zongliang, Harwig Alex, Berkhout Ben, Herrera-Carrillo Elena

机构信息

a Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and immunity Amsterdam (CINIMA), Academic Medical Center , University of Amsterdam , Amsterdam , The Netherlands.

出版信息

Transcription. 2017;8(5):275-287. doi: 10.1080/21541264.2017.1322170. Epub 2017 Jun 9.

DOI:10.1080/21541264.2017.1322170
PMID:28598252
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5703244/
Abstract

Type 3 RNA polymerase III (Pol III) promoters are widely used for the expression of small RNAs such as short hairpin RNA and guide RNA in the popular RNAi and CRISPR-Cas gene regulation systems. Although it is generally believed that type 3 Pol III promoters use a defined transcription start site (+1 position), most man-made promoter constructs contain local sequence alterations of which the impact on transcription efficiency and initiation accuracy is not known. For three human type 3 Pol III promoters (7SK, U6, and H1), we demonstrated that the nucleotides around the +1 position affect both the transcriptional efficiency and start site selection. Human 7SK and U6 promoters with A or G at the +1 position efficiently produced small RNAs with a precise +1 start site. The human H1 promoter with +1A or G also efficiently produced small RNAs but from multiple start sites in the -3/-1 window. These results provide new insights for the design of vectors for accurate expression of designed small RNAs for research and therapeutic purposes.

摘要

3型RNA聚合酶III（Pol III）启动子被广泛用于在流行的RNA干扰（RNAi）和CRISPR-Cas基因调控系统中表达小RNA，如短发夹RNA和引导RNA。虽然人们普遍认为3型Pol III启动子使用一个确定的转录起始位点（+1位置），但大多数人工启动子构建体都包含局部序列改变，其对转录效率和起始准确性的影响尚不清楚。对于三种人类3型Pol III启动子（7SK、U6和H1），我们证明了+1位置周围的核苷酸会影响转录效率和起始位点选择。在+1位置带有A或G的人类7SK和U6启动子能高效产生具有精确+1起始位点的小RNA。带有+1A或G的人类H1启动子也能高效产生小RNA，但来自-3/-1窗口中的多个起始位点。这些结果为设计用于准确表达用于研究和治疗目的的设计小RNA的载体提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/8e7a1739336f/ktrn-08-05-1322170-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/97eb571648d5/ktrn-08-05-1322170-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/e163bf3333ea/ktrn-08-05-1322170-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/1a5e13078b5b/ktrn-08-05-1322170-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/83e9ab823699/ktrn-08-05-1322170-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/4fdbc8e9ca6d/ktrn-08-05-1322170-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/7308ef30d044/ktrn-08-05-1322170-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/d4e885033379/ktrn-08-05-1322170-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/8e7a1739336f/ktrn-08-05-1322170-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/97eb571648d5/ktrn-08-05-1322170-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/e163bf3333ea/ktrn-08-05-1322170-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/1a5e13078b5b/ktrn-08-05-1322170-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/83e9ab823699/ktrn-08-05-1322170-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/4fdbc8e9ca6d/ktrn-08-05-1322170-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/7308ef30d044/ktrn-08-05-1322170-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/d4e885033379/ktrn-08-05-1322170-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b36/5703244/8e7a1739336f/ktrn-08-05-1322170-g008.jpg

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