title: A non-surgical approach for male germ cell mediated gene transmission through transgenesis.
abstract: Microinjection of foreign DNA in male pronucleus by in-vitro embryo manipulation is difficult but remains the method of choice for generating transgenic animals. Other procedures, including retroviral and embryonic stem cell mediated transgenesis are equally complicated and have limitations. Although our previously reported technique of testicular transgenesis circumvented several limitations, it involved many steps, including surgery and hemicastration, which carried risk of infection and impotency. We improved this technique further, into a two step non-surgical electroporation procedure, for making transgenic mice. In this approach, transgene was delivered inside both testes by injection and modified parameters of electroporation were used for in-vivo gene integration in germ cells. Using variety of constructs, germ cell integration of the gene and its transmission in progeny was confirmed by PCR, slot blot and immunohistochemical analysis. This improved technique is efficient, requires substantially less time and can be easily adopted by various biomedical researchers.
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India.
authors.forename: Abul
authors.initials: A
authors.lastname: Usmani
authors.forename: Nirmalya
authors.initials: N
authors.lastname: Ganguli
authors.forename: Hironmoy
authors.initials: H
authors.lastname: Sarkar
authors.forename: Suveera
authors.initials: S
authors.lastname: Dhup
authors.forename: Suryaprakash R
authors.initials: SR
authors.lastname: Batta
authors.forename: Manoj
authors.initials: M
authors.lastname: Vimal
authors.forename: Nilanjana
authors.initials: N
authors.lastname: Ganguli
authors.forename: Sayon
authors.initials: S
authors.lastname: Basu
authors.forename: P
authors.initials: P
authors.lastname: Nagarajan
authors.forename: Subeer S
authors.initials: SS
authors.lastname: Majumdar
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D018274:Electroporation / Q000379:methods; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005854:Germ Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D013737:Testis / Q000378:metabolism; D019076:Transgenes