title: Genome editing in mouse spermatogonial stem/progenitor cells using engineered nucleases.
abstract: Editing the genome to create specific sequence modifications is a powerful way to study gene function and promises future applicability to gene therapy. Creation of precise modifications requires homologous recombination, a very rare event in most cell types that can be stimulated by introducing a double strand break near the target sequence. One method to create a double strand break in a particular sequence is with a custom designed nuclease. We used engineered nucleases to stimulate homologous recombination to correct a mutant gene in mouse "GS" (germline stem) cells, testicular derived cell cultures containing spermatogonial stem cells and progenitor cells. We demonstrated that gene-corrected cells maintained several properties of spermatogonial stem/progenitor cells including the ability to colonize following testicular transplantation. This proof of concept for genome editing in GS cells impacts both cell therapy and basic research given the potential for GS cells to be propagated in vitro, contribute to the germline in vivo following testicular transplantation or become reprogrammed to pluripotency in vitro.
authors.affiliation: Department of Chemistry, Indiana University, Bloomington, Indiana, United States of America.
authors.forename: Danielle A
authors.initials: DA
authors.lastname: Fanslow
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, United States of America.
authors.forename: Stacey E
authors.initials: SE
authors.lastname: Wirt
authors.affiliation: Department of Biochemistry, UT Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Jenny C
authors.initials: JC
authors.lastname: Barker
authors.affiliation: Department of Biochemistry, UT Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Jon P
authors.initials: JP
authors.lastname: Connelly
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, California, United States of America.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Porteus
authors.affiliation: Department of Chemistry, Indiana University, Bloomington, Indiana, United States of America.
authors.forename: Christina Tenenhaus
authors.initials: CT
authors.lastname: Dann
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism* / Q000637:transplantation; D000818:Animals; D002460:Cell Line; D004720:Endonucleases / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D008297:Male; D051379:Mice; D010641:Phenotype; D013094:Spermatozoa / Q000166:cytology; D033581:Stem Cell Transplantation; D013737:Testis / Q000166:cytology
chemical_list: D004720:Endonucleases