| Autor: |
Purdy AK; Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA, USA., Campbell KS |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2010; Vol. 612, pp. 223-31. |
| DOI: |
10.1007/978-1-60761-362-6_15 |
| Abstrakt: |
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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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