Selected technologies to control genes and their products for experimental and clinical purposes.
"On-demand" regulation of gene expression is a powerful tool to elucidate the functions of proteins and biologically-active RNAs. We describe here three different approaches to the regulation of expression or activity of genes or proteins. Promoter-based regulation of gene expression was among the most rapidly developing techniques in the 1980s and 1990 s. Here we provide basic information and also some characteristics of the metallothionein-promoter-based system, the tet-off system, Muristerone-A-regulated expression through the ecdysone response element, RheoSwitch, coumermycin/novobiocin-regulated gene expression, chemical dimerizer-based promoter activation systems, the "Dual Drug Control" system, "constitutive androstane receptor"-based regulation of gene expression, and RU486/mifepristone-driven regulation of promoter activity. A large part of the review concentrates on the principles and usage of various RNA interference techniques (RNAi: siRNA, shRNA, and miRNA-based methods). Finally, the last part of the review deals with historically the oldest, but still widely used, methods of temperature-dependent regulation of enzymatic activity or protein stability (temperature-sensitive mutants). Due to space limitations we do not describe in detail but just mention the tet-regulated systems and also fusion-protein-based regulation of protein activity, such as estrogen-receptor fusion proteins. The information provided below is aimed to assist researchers in choosing the most appropriate method for the planned development of experimental systems with regulated expression or activity of studied proteins.
Alexander
Helen K
HK
Cancer Care Manitoba, Manitoba Institute of Cell Biology, Winnipeg, Canada.
Booy
Evan P
EP
Xiao
Wenyan
W
Ezzati
Peyman
P
Baust
Heinrich
H
Los
Marek
M
D000818:Animals; D005786:Gene Expression Regulation; D005821:Genetic Techniques; D050505:Mutant Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D013696:Temperature
D050505:Mutant Proteins