RNA interference in immune cells by use of osmotic delivery of siRNA.
Delivery of siRNA to immune cells has been one of the major obstacles to widespread application of RNAi in the immunology field. Here, we report that osmotic delivery of siRNA can be used to silence genes in macrophage RAW264.7 without incurring either cytotoxic or immunomodulatory activity. We also showed usefulness of the osmotic delivery in other types of cells including T cell DO11.10. By repeated osmotic delivery of siRNA, long-term gene silencing was readily achieved. When TLR4 was disrupted in RAW264.7 cells for 48 h and the cells were stimulated with the TLR4 ligand LPS, a significant decrease in TNFalpha production was observed. DNA microarray-based gene expression profile analysis showed that gene silencing by osmotic delivery of siRNA was target-specific and the delivery method itself had little influence on overall gene expression.
Aoki
Mikio
M
Genomic Science Laboratories, Dainippon Sumitomo Pharma Co., Ltd., 3-1-98 Kasudade Naka, Konohana-ku, Osaka 554-0022, Japan.
Ishii
Takayuki
T
Kanaoka
Masaharu
M
Kimura
Toru
T
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003609:Dactinomycin / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D007107:Immune System; D008070:Lipopolysaccharides / Q000737:chemistry; D008264:Macrophages; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D009995:Osmosis; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D013997:Time Factors; D051193:Toll-Like Receptors / Q000378:metabolism; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D018840:HSP70 Heat-Shock Proteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; C081884:heat-shock protein 70.1; D003609:Dactinomycin; C025942:7-aminoactinomycin D; D016650:Fluorescein-5-isothiocyanate