Document Detail


Construction of shRNA Expression Plasmids for Silkworm Cell Lines Using Single-Stranded DNA and Bst DNA Polymerase.
MedLine Citation:
PMID:  23027060     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Transfection of short hairpin RNA (shRNA) expression plasmids is conventionally performed for gene-specific knockdown in cultured mammalian and insect cells. Here, I describe a simple method to synthesize an inverted repeat DNA in a U6 small nuclear RNA promoter-based parent vector using a single-stranded inverted repeat DNA and Bst DNA polymerase. The shRNA expression plasmids constructed by this method were confirmed to promote efficient RNA interference knockdown in silkworm cell lines. This method may be useful for constructing a relatively large number of shRNA expression plasmids.
Authors:
Hiromitsu Tanaka
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  942     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-10-02     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  347-55     Citation Subset:  IM    
Affiliation:
Insect Mimetics Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan, htanaka1@affrc.go.jp.
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