This is an archive of papers published by the staff and faculty of Fox Chase Cancer Center. For questions about content, please contact Talbot Research Library
Last updated on
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
AbstractNatural 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.
NotesPurdy, 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.