title: A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice.
abstract: Lineage tracing experiments define the origin, fate and behavior of cells in a specific tissue or organism. This technique has been successfully applied for many decades, revealing seminal findings in developmental biology. More recently, it was adopted by stem cell biologists to identify and track different stem cell populations with minimal experimental intervention. The recent developments in mouse genetics, the availability of a large number of mouse strains, and the advancements in fluorescent microscopy allow the straightforward design of powerful lineage tracing systems for various tissues with basic expertise, using commercially available tools. We have recently taken advantage of this powerful methodology to explore the origin and fate of stem cells at the ocular surface using R26R-Confetti mouse. This model offers a multi-color genetic system, for the expression of 4 fluorescent genes in a random manner. Here we describe the principles of this methodology and provide an adaptable protocol for designing lineage tracing experiments; specifically for the corneal epithelium as well as for other tissues.
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology.
authors.forename: Aya
authors.initials: A
authors.lastname: Amitai-Lange
authors.affiliation: Department of Ophthalmology, Hillel Yaffe Medical Center.
authors.forename: Eran
authors.initials: E
authors.lastname: Berkowitz
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology.
authors.forename: Anna
authors.initials: A
authors.lastname: Altshuler
authors.affiliation: Department of Ophthalmology, Hillel Yaffe Medical Center.
authors.forename: Noora
authors.initials: N
authors.lastname: Dbayat
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology.
authors.forename: Waseem
authors.initials: W
authors.lastname: Nasser
authors.affiliation: Bioimging Center, Biomedical Core Facility, The Ruth and Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology.
authors.forename: Edith
authors.initials: E
authors.lastname: Suss-Toby
authors.affiliation: Department of Ophthalmology, Hillel Yaffe Medical Center.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Tiosano
authors.affiliation: Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, Technion, Israel Institute of Technology; shalomfe@tx.technion.ac.il.
authors.forename: Ruby
authors.initials: R
authors.lastname: Shalom-Feuerstein
mesh_terms: D000483:Alleles; D000818:Animals; D019070:Cell Lineage; D003315:Cornea / Q000166:cytology*; D017930:Genes, Reporter; D019426:Integrases / Q000096:biosynthesis / Q000235:genetics; D008164:Luminescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000379:methods; D013050:Spectrometry, Fluorescence / Q000379:methods; D013234:Stem Cells / Q000166:cytology*
chemical_list: D008164:Luminescent Proteins; C045073:Cre recombinase; D019426:Integrases