Post-transcriptional gene silencing induced by short interfering RNAs in cultured transgenic plant cells.
Short interfering RNA (siRNA) is widely used for studying post-transcriptional gene silencing and holds great promise as a tool for both identifying function of novel genes and validating drug targets. Two siRNA fragments (siRNA-a and -b), which were designed against different specific areas of coding region of the same target green fluorescent protein (GFP) gene, were used to silence GFP expression in cultured gfp transgenic cells of rice (Oryza sativa L.; OS), cotton (Gossypium hirsutum L.; GH), Fraser fir [Abies fraseri (Pursh) Poir; AF], and Virginia pine (Pinus virginiana Mill.; PV). Differential gene silencing was observed in the bombarded transgenic cells between two siRNAs, and these results were consistent with the inactivation of GFP confirmed by laser scanning microscopy, Northern blot, and siRNA analysis in tested transgenic cell cultures. These data suggest that siRNA-mediated gene inactivation can be the siRNA specific in different plant species. These results indicate that siRNA is a highly specific tool for targeted gene knockdown and for establishing siRNA-mediated gene silencing, which could be a reliable approach for large-scale screening of gene function and drug target validation.
Tang
Wei
W
Department of Biology, Howell Science Complex, East Carolina University, Greenville, NC 27858-4353, USA. tangw@mail.ecu.edu
Samuels
Vanessa
V
Whitley
Nicki
N
Bloom
Nicole
N
DeLaGarza
Tinya
T
Newton
Ronald J
RJ
D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010944:Plants / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000378:metabolism*; D013997:Time Factors
D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins