title: Generation of mouse lines conditionally over-expressing microRNA using the Rosa26-Lox-Stop-Lox system.
abstract: MicroRNAs are currently the object of intensive investigation due to their role in a myriad of physiological processes and pathological conditions, such as gene regulation and tumorigenesis. To better understand microRNA function, numerous laboratories have already taken advantage of the available techniques of genome editing in mouse. Here, we describe how to generate genetically engineered mouse lines using the popular Rosa-26 Lox-Stop-Lox Knock-In (Rosa-LSL-KI) targeting. This strategy allows for the selective overexpression of microRNAs of interest when coupled to a tissue-specific Cre-expressing line. The present protocol illustrates in detail both the engineering of the targeting vector and the generation of mutated ES clones ready for injection into mouse blastocysts.
authors: Department of Molecular Virology, Immunology and Medical Genetics, Wexner Medical Center, Comprehensive Cancer Center, The Ohio State University, Biomedical Research Tower, Room 988, 460W. 12th Avenue, Columbus, OH, 43210, USA.
authors: Claudia
authors: C
authors: Piovan
authors: Foued
authors: F
authors: Amari
authors: Francesca
authors: F
authors: Lovat
authors: Qun
authors: Q
authors: Chen
authors: Vincenzo
authors: V
authors: Coppola
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D002460:Cell Line; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism; D015925:Cryopreservation; D018274:Electroporation; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D055879:Gene Knock-In Techniques; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D019426:Integrases / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D011247:Pregnancy
chemical_list: D035683:MicroRNAs; C045073:Cre recombinase; D019426:Integrases