NDLI: Regulation of U6 promoter activity by transcriptional interference in viral vector-based RNAi.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Nie, Linghu Das Thakur, Meghna Wang, Yumei Su, Qin Zhao, Yongliang Feng, Yunfeng
Description	Country affiliation: China Author Affiliation: Nie L ( Key Laboratory of Genome Sciences and Information, Cancer Biology and Genetics Group, Beijing Institute of Genomics, Chinese Acadamy of Sciences, Beijing 100029, China.)
Abstract	The direct negative impact of the transcriptional activity of one component on the second one in cis is referred to as transcriptional interference (TI). U6 is a type III RNA polymerase III promoter commonly used for driving small hairpin RNA (shRNA) expression in vector-based RNAi. In the design and construction of viral vectors, multiple transcription units may be arranged in close proximity in a space-limited vector. Determining if U6 promoter activity can be affected by TI is critical for the expression of target shRNA in gene therapy or loss-of-function studies. In this research, we designed and implemented a modified retroviral system where shRNA and exogenous gene expressions were driven by two independent transcriptional units. We arranged U6 promoter driving shRNA expression and UbiC promoter in two promoter arrangements. In primary macrophages, we found U6 promoter activity was inhibited by UbiC promoter when in the divergent arrangement but not in tandem. In contrast, PKG promoter had no such negative impact. Instead of enhancing U6 promoter activity, CMV enhancer had significant negative impact on U6 promoter activity in the presence of UbiC promoter. Our results indicate that U6 promoter activity can be affected by TI in a proximal promoter-specific and arrangement-dependent manner.
File Format	HTM / HTML
ISSN	16720229
Issue Number	3
Volume Number	8
e-ISSN	22103244
Journal	Genomics, Proteomics & Bioinformatics
Language	English
Publisher	Elsevier
Publisher Date	2010-09-01
Publisher Place	China
Access Restriction	Subscribed
Subject Keyword	Discipline Genetics Discipline Biochemistry Discipline Computational Biology Genetic Vectors Genetics Promoter Regions, Genetic Rna Interference Rna, Small Nuclear Transcription, Genetic Animals Cytomegalovirus Genetic Techniques Mice Models, Genetic Oxo-acid-lyases Polymerase Chain Reaction Methods Rna, Small Interfering Metabolism Retroviridae Journal Article
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology Biochemistry Computational Mathematics

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