Introduction of shRNAs into human NK-like cell lines with retrovirus.
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.
Purdy
Amanda K
AK
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA, USA.
Campbell
Kerry S
KS
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D055029:Inverted Repeat Sequences; D007694:Killer Cells, Natural / Q000166:cytology / Q000378:metabolism*; D008969:Molecular Sequence Data; D012313:RNA / Q000235:genetics* / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection
D012313:RNA