TY - JOUR
T1 - Introduction of shRNAs into human NK-like cell lines with retrovirus.
AU - Purdy, Amanda K.
AU - Campbell, Kerry S.
PY - 2010
Y1 - 2010
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=77449104490&partnerID=8YFLogxK
U2 - 10.1007/978-1-60761-362-6_15
DO - 10.1007/978-1-60761-362-6_15
M3 - Article
C2 - 20033644
AN - SCOPUS:77449104490
SN - 1064-3745
VL - 612
SP - 223
EP - 231
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -