TY - JOUR
T1 - Ribozyme- and siRNA-mediated suppression of RGS-containing RhoGEF proteins
AU - Wang, Qin
AU - Liu, Min
AU - Kozasa, Tohru
AU - Rothestein, Jeffrey D.
AU - Sternweis, Paul C.
AU - Neubig, Richard R.
N1 - Funding Information:
This work was supported by NIH Grant GM39561 awarded to R.R.N.
PY - 2004
Y1 - 2004
N2 - 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.
AB - 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.
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U2 - 10.1016/S0076-6879(04)89015-3
DO - 10.1016/S0076-6879(04)89015-3
M3 - Article
C2 - 15313570
AN - SCOPUS:4344609075
SN - 0076-6879
VL - 389
SP - 244
EP - 265
JO - Methods in Enzymology
JF - Methods in Enzymology
ER -