title: Lentiviral mediated transgenesis by in vivo manipulation of spermatogonial stem cells.
abstract: This report describes a technique for the generation of transgenic mice by in vivo manipulation of spermatogonial stem cells with a high rate of success. Spermatogonial stem cells (SSCs) in pre-pubescent animals were infected in vivo with recombinant lentiviruses expressing EGFP-f and mated with normal females. All male pre-founder mice produced transgenic pups with an overall success rate of over 60%. The transgene was heritable and the pre-founder mice could be used in multiple mating experiments. This technology could be used to perform overexpression/knockdown screens in vivo using bar-coded lentiviruses, thus permitting the design of genetic screens in the mouse. Further, this technology could be adapted to other laboratory animals resulting in the generation of model systems that closely approximate human development and disease.
authors.affiliation: KS-215, Advanced Centre for Treatment Research Education and Cancer, Tata Memorial Centre, Kharghar Node, Navi Mumbai, India.
authors.forename: Lalit
authors.initials: L
authors.lastname: Sehgal
authors.forename: Rahul
authors.initials: R
authors.lastname: Thorat
authors.forename: Nileema
authors.initials: N
authors.lastname: Khapare
authors.forename: Amitabha
authors.initials: A
authors.lastname: Mukhopadhaya
authors.forename: Mugdha
authors.initials: M
authors.lastname: Sawant
authors.forename: Sorab N
authors.initials: SN
authors.lastname: Dalal
mesh_terms: D000818:Animals; D003433:Crosses, Genetic; D005260:Female; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D016254:Mutagenesis, Insertional / Q000235:genetics; D013093:Spermatogonia / Q000166:cytology* / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins