PT  - JOURNAL ARTICLE
AU  - Abul Usmani
AU  - Nirmalya Ganguli
AU  - Subodh K Jain
AU  - Nilanjana Ganguli
AU  - Rajesh Kumar Sarkar
AU  - Mayank Choubey
AU  - Mansi Shukla
AU  - Hironmoy Sarkar
AU  - Subeer S Majumdar
TI  - Robust Generation of Transgenic Mice by Hypotonic Shock Mediated Transgene Delivery in Testicular Germ Cells
AID  - 10.1101/049239
DP  - 2016 Jan 01
TA  - bioRxiv
PG  - 049239
4099  - http://biorxiv.org/content/early/2016/06/16/049239.short
4100  - http://biorxiv.org/content/early/2016/06/16/049239.full
AB  - 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.