RT Journal Article
SR Electronic
T1 Robust Generation of Transgenic Mice by Hypotonic Shock Mediated Transgene Delivery in Testicular Germ Cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 049239
DO 10.1101/049239
A1 Abul Usmani
A1 Nirmalya Ganguli
A1 Subodh K Jain
A1 Nilanjana Ganguli
A1 Rajesh Kumar Sarkar
A1 Mayank Choubey
A1 Mansi Shukla
A1 Hironmoy Sarkar
A1 Subeer S Majumdar
YR 2016
UL http://biorxiv.org/content/early/2016/06/16/049239.abstract
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.