Identification and characterization of transcription factor target genes using gene-targeted mice.
Critical to understanding biological roles for transcription factors is an appreciation of the target genes regulated by the transcription factor. The identification of target genes can often expand the understanding of known biological roles for a transcription factor and may reveal unappreciated and unexpected functions. This article focuses on the identification and characterization of transcription factor target genes using mouse molecular genetics. The use of genetically engineered (knockout) mice and global gene expression analysis to identify transcription factor target genes is reviewed, with emphasis on important technical considerations. Detailed protocols for the application of real-time reverse transcription polymerase chain reaction and immunohistochemistry in target gene expression analysis are described. The article closes with a discussion of the use of mouse molecular genetics for the characterization of target genes as downstream effectors of transcription factors.
DeRyckere
Deborah
D
Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, BRB802, 4200 East Ninth Avenue, Denver, CO 80262, USA.
DeGregori
James
J
D000465:Algorithms; D000818:Animals; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D020869:Gene Expression Profiling; D018390:Gene Targeting; D018398:Homeodomain Proteins / Q000235:genetics; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D012160:Retina / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D018398:Homeodomain Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C109288:cone rod homeobox protein; D012313:RNA