Reporter genes for monitoring gene expression in mammalian cells

摘要: Publisher Summary The advent of reporter gene technology has greatly facilitated the knowledge mechanisms many cellular processes Reporter genes encode proteins with phenotypic properties that are both distinct (from system being studied) and conveniently monitored. Linkage activity assayed to expression is accomplished by fusion appropriate DNA sequences gene. These either regulatory in nature, typically responsive event under examination, or structural, encoding mediate such an event. After introduction chimera into cells, qualitative quantitative extrapolated from linked product. Application reporter–gene mammalian cells was introduced 1982 development plasmid vectors, bacterial enzymes chloramphenicol acetyltransferase (CAT) β-galactosidase (β-gal), study eukaryotic regulation. As these initial studies, have been traditionally used characterize dissect transcriptionally active regions. Although characterization cis-elements remains a common usage, attained increasing levels sophistication now numerous other applications: transcription factors associated as co-activators, delineation signal transduction pathways, identification protein–protein interactions, determination cell fates, visualization trafficking, high-throughput screening chemicals for adverse therapeutic effects, optimization monitoring delivery systems. Numerous genes, prokaryotic origin, proposed use cells. Only handful these, however, widely this capacity. Aside obvious characteristic products cannot be toxic, commonly reporters share two features: (1) exhibit characteristics unique (i.e., foreign cells) can easily distinguished any similar endogenous activity, (2) methods detection sensitive, generally quantitative, broad linear dynamic range; they easy perform, reproducible reasonably cost-effective. In terms themselves, single most significant since time evolution green fluorescent protein (GFP) powerful multi-faceted protein. chapter discusses well some