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Development of siRNA systems to knock‐down arginase I‐specific or iNOS‐specific gene expression
Author(s) -
Guo Zhong,
Bucher Brian T,
Shao Lifang,
Geller David A
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1120.1
Subject(s) - arginase , small interfering rna , transfection , biology , gene expression , microbiology and biotechnology , gene , arginine , biochemistry , amino acid
The L‐arginine/iNOS pathway has been recognized to play critical roles during infection, inflammation, organ injury, and transplant rejection. The development of vector‐based siRNAs provides a powerful genetic approach to study the pathophysiology in vitro and in vivo by suppressing the endogenous arginase I or iNOS expression. Here, we have screened and identified the siRNA sequences specific for arginase I or iNOS gene, which are both conserved among human, rat and mouse species. Transfection of RAW cells with arginase I‐specific siRNA significantly reduces arginase‐I protein expression and arginase activity after stimulation with cytokines. Moreover, Transfection of DLD −1 cells with iNOS‐specific siRNA also significantly decreased iNOS protein expression and nitrite production after stimulation with cytokines. Non‐specific siRNA served as control, and did not knock‐down either gene expression. These siRNAs should provide useful tools to study the effects of iNOS and arginase I expression in biologic systems.