title: Robust generation of transgenic mice by simple hypotonic solution mediated delivery of transgene in testicular germ cells.
abstract: Our ability to decipher gene sequences has increased enormously with the advent of modern sequencing tools, but the ability to divulge functions of new genes have not increased correspondingly. This has caused a remarkable delay in functional interpretation of several newly found genes in tissue and age specific manner, limiting the pace of biological research. This is mainly due to lack of advancements in methodological tools for transgenesis. Predominantly practiced method of transgenesis by pronuclear DNA-microinjection is time consuming, tedious, and requires highly skilled persons for embryo-manipulation. Testicular electroporation mediated transgenesis requires use of electric current to testis. To this end, we have now developed an innovative technique for making transgenic mice by giving hypotonic shock to male germ cells for the gene delivery. Desired transgene was suspended in hypotonic Tris-HCl solution (pH 7.0) and simply injected in testis. This resulted in internalization of the transgene in dividing germ-cells residing at basal compartment of tubules leading to its integration in native genome of mice. Such males generated transgenic progeny by natural mating. Several transgenic animals can be generated with minimum skill within short span of time by this easily adaptable novel technique.
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India; Department of Zoology, Dr. H. S. Gour University, Sagar, India.
authors.forename: Abul
authors.initials: A
authors.lastname: Usmani
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Nirmalya
authors.initials: N
authors.lastname: Ganguli
authors.affiliation: Department of Zoology, Dr. H. S. Gour University , Sagar, India.
authors.forename: Subodh K
authors.initials: SK
authors.lastname: Jain
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Nilanjana
authors.initials: N
authors.lastname: Ganguli
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Rajesh Kumar
authors.initials: RK
authors.lastname: Sarkar
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Mayank
authors.initials: M
authors.lastname: Choubey
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Mansi
authors.initials: M
authors.lastname: Shukla
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology , New Delhi, India.
authors.forename: Hironmoy
authors.initials: H
authors.lastname: Sarkar
authors.affiliation: Embryo Biotechnology Laboratory, National Institute of Immunology, New Delhi, India; National Institute of Animal Biotechnology, Hyderabad, Telangana, India.
authors.forename: Subeer S
authors.initials: SS
authors.lastname: Majumdar