Small interfering RNA-mediated gene silencing in T lymphocytes.
Introduction of small interfering RNAs (siRNAs) into a cell can cause a specific interference of gene expression known as RNA interference (RNAi). However, RNAi activity in lymphocytes and in normal primary mammalian cells has not been thoroughly demonstrated. In this report, we show that siRNAs complementary to CD4 and CD8alpha specifically reduce surface expression of these coreceptors and their respective mRNA in a thymoma cell line model. We show that RNAi activity is only caused by a subset of siRNAs complementary to the mRNA target and that ineffective siRNAs can compete with effective siRNAs. Using primary differentiated T lymphocytes, we provide the first evidence of siRNA-mediated RNAi gene silencing in normal nontransformed somatic mammalian lymphocytes.
McManus
Michael T
MT
Center for Cancer Research, Massachusetts Institute of Technology, 40 Ames Street E17-526, Cambridge, MA 02139, USA.
Haines
Brian B
BB
Dillon
Christopher P
CP
Whitehurst
Charles E
CE
van Parijs
Luk
L
Chen
Jianzhu
J
Sharp
Phillip A
PA
D000818:Animals; D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics; D016827:CD8 Antigens / Q000096:biosynthesis / Q000235:genetics; D020868:Gene Silencing / Q000276:immunology*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D020871:RNA Stability / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry*; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured
D015704:CD4 Antigens; D016827:CD8 Antigens; C431698:CD8alpha antigen; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins