Savan Ram, Chan Tim, Young Howard A
Laboratory of Experimental Immunology, Cancer and Inflammation Program, National Institutes of Health-Frederick, Frederick, MD, USA.
Methods Mol Biol. 2010;612:209-21. doi: 10.1007/978-1-60761-362-6_14.
Natural killer (NK) cells play a vital role in the control of cancer and microbial infections. A major hinderance in studying NK cells is the resistance of these cells to gene transfer. Considering over-expression and gene knockdown studies are crucial tools to study the biology of cells, technologies suitable for transferring genes into NK cells are invaluable. Among various technologies available for gene transfer, lentiviral-mediated transduction has been successful in introducing genes into NK cells. We have standardized methods of lentiviral infection in human and mouse NK cell lines. We obtain transduction efficiencies of 15% in the NK-92 cell line and 30-40% in LNK, YT, and DERL7 cell lines. This method allows efficient and stable introduction of genes and shRNAs into NK cell lines.
自然杀伤（NK）细胞在癌症控制和微生物感染中发挥着至关重要的作用。研究NK细胞的一个主要障碍是这些细胞对基因转移具有抗性。鉴于过表达和基因敲低研究是研究细胞生物学的关键工具，适用于将基因转入NK细胞的技术非常宝贵。在可用于基因转移的各种技术中，慢病毒介导的转导已成功地将基因导入NK细胞。我们已经标准化了在人和小鼠NK细胞系中进行慢病毒感染的方法。我们在NK-92细胞系中获得了15%的转导效率，在LNK、YT和DERL7细胞系中获得了30-40%的转导效率。该方法能够高效、稳定地将基因和短发夹RNA（shRNA）导入NK细胞系。
