Mouse models for miRNA expression: the ROSA26 locus.
In 1991, Soriano and coworkers isolated the ROSA26 locus in a gene-trap mutagenesis screening performed in mouse embryonic stem (ES) cells. The ubiquitous expression of ROSA26 in embryonic and adult tissues, together with the high frequency of gene-targeting events observed at this locus in murine ES cells has led to the establishment in the past 10 years of over 130 knock-in lines expressing successfully from the ROSA26 locus a variety of transgenes including reporters, site-specific recombinases and, recently, noncoding RNAs. Different strategies can be employed to drive transgene expression from the ROSA26 locus. This chapter provides an overview of the current methodologies used to generate ROSA26 knock-in lines and describes different approaches that exploit the ROSA26 gene to control expression of transgenes, including miRNAs, in a temporal, cell-type, and stage-specific fashion.
Casola
Stefano
S
IFOM, The FIRC Institute of Molecular Oncology Foundation, Milan, Italy.
D000818:Animals; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques / Q000295:instrumentation / Q000379:methods; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D022661:RNA, Untranslated; D011995:Recombination, Genetic; D014162:Transfection / Q000295:instrumentation / Q000379:methods; D019076:Transgenes
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D035683:MicroRNAs; D011506:Proteins; D022661:RNA, Untranslated