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Purdy AK , Campbell KS
Introduction of shRNAs into human NK-like cell lines with retrovirus
Methods Mol Biol. 2010 ;612 :223-31
PMID: 20033644    PMCID: PMC2798137   
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Abstract
Natural killer (NK) cell lines are difficult to transfect using standard techniques, which limits the ability to establish long-term knockdown of proteins with short-hairpin (sh)RNAs. We have developed a method to stably knockdown protein expression in human NK-like lines by introducing shRNAs in retroviral vectors. After a single transduction with retrovirus, shRNA-containing cells can be selected with drug treatment or sorted for enhanced green fluorescent protein (EGFP) expression. With this method, protein expression can be stably decreased to less than 10% of wild-type levels.
Notes
Purdy, Amanda K Campbell, Kerry S CA06927/CA/NCI NIH HHS/United States R01-CA083859/CA/NCI NIH HHS/United States R01-CA100226/CA/NCI NIH HHS/United States T32-CA009035/CA/NCI NIH HHS/United States Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States Methods in molecular biology (Clifton, N.J.) Nihms152767 Methods Mol Biol. 2010;612:223-31.