title: A conditional mouse line for lineage tracing of Sox9 loss-of-function cells using enhanced green fluorescent protein.
abstract: Traditionally, conditional knockout studies in mouse have utilized the Cre or Flpe technology to activate the expression of reporter genes such as lacZ or PLAP. Employing these reporter genes, however, requires tissue fixation. To make way for downstream in vivo or in vitro applications, we have inserted enhanced green fluorescent protein (EGFP) into the endogenous Sox9 locus and generated a novel conditional Sox9 null allele, by flanking the entire Sox9 coding region with loxP sites and inserting an EGFP reporter gene into the 3'-UTR allowing for EGFP to be expressed upon Sox9 loss of function yet under the control of the endogenous Sox9 promoter. Mating this new allele to any Cre-expressing line, the fate of Sox9 null cells can be traced in the cell type of interest in vivo or in vitro after fluorescence-activated cell sorting.
authors.affiliation: Stem Cell and Developmental Biology Department, Genome Institute of Singapore, 60 Biopolis Street, Singapore, 138672, Singapore, chatterjees@gis.a-star.edu.sg.
authors.forename: Sumantra
authors.initials: S
authors.lastname: Chatterjee
authors.forename: Petra
authors.initials: P
authors.lastname: Kraus
authors.forename: V
authors.initials: V
authors.lastname: Sivakamasundari
authors.forename: Xing
authors.initials: X
authors.lastname: Xing
authors.forename: Sook Peng
authors.initials: SP
authors.lastname: Yap
authors.forename: Song
authors.initials: S
authors.lastname: Jie
authors.forename: Thomas
authors.initials: T
authors.lastname: Lufkin
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D055758:SOX9 Transcription Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: C447413:Col2a1 protein, mouse; D024043:Collagen Type II; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins