Ribozyme- and siRNA-mediated suppression of RGS-containing RhoGEF proteins.
Given recent efforts to determine the sequence information on thousands of genes in the human genome, the current challenge is to identify the functions of these genes, including those encoding the regulator of G-protein signaling protein gene superfamily, and to establish their roles in particular signaling pathways in a native system. Increasingly, reverse genetic approaches are being used to address these questions. This article compares two powerful approaches [ribozyme and "short interfering" RNA (siRNA) techniques] under identical conditions for the first report on the suppression of endogenous RGS domain-containing RhoGEFs. The siRNA technique was found to be much more potent than ribozyme targeting at the same mRNA site of RGS-RhoGEFs. Also, the three siRNAs targeting LARG, PDZ-RhoGEF, and p115-RhoGEF are able to discriminate the closely related sequences within this RGS-RhoGEF gene family.
Wang
Qin
Q
Department of Pharmacology, University of Michigan, Ann Arbor 48109, USA.
Liu
Min
M
Kozasa
Tohru
T
Rothestein
Jeffrey D
JD
Sternweis
Paul C
PC
Neubig
Richard R
RR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D017434:Protein Structure, Tertiary; D020710:RGS Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D034622:RNA Interference; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020662:Guanine Nucleotide Exchange Factors; D020710:RGS Proteins; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D008156:Luciferases