@article {Usmani049239, author = {Abul Usmani and Nirmalya Ganguli and Subodh K Jain and Nilanjana Ganguli and Rajesh Kumar Sarkar and Mayank Choubey and Hironmoy Sarkar and Subeer S Majumdar}, title = {Robust generation of transgenic mice by hypotonic shock mediated transgene delivery in testicular germ cells}, elocation-id = {049239}, year = {2016}, doi = {10.1101/049239}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Although our ability to decipher gene sequences has increased enormously with the advent of modern sequencing tools, the efficiency of obtaining information about functions of new genes have not increased correspondingly. This is mainly due to lack of advancements in methods of transgenesis. Presently practiced zygotic pronuclear DNA-microinjection is time consuming, tedious and requires highly skilled persons for embryo-manipulation. Hence, we have established an innovative technique for making transgenic mice by hypotonic-shock driven gene delivery in male germ cells. For this, suspension of transgene in hypotonic Tris-HCl solution was delivered in testis non-surgically by simple injection. This resulted in internalization and integration of the transgene in dividing germ-cells by hypotonic-shock leading to generation of transgenic progeny by such males. We have also achieved gene silencing by using construct containing shRNA for specific mRNA. Several transgenic animals can be generated within short span of time by this easily adaptable novel technique.Method Summary We present here a novel method for generating transgenic mice by non-surgical in vivo transfection of testis. Transfection is achieved by simple injection of testis with transgene suspended in hypotonic solution of Tris-HCl. We show here transgene is stably transferred in progenies generated from transfected male mice. By this new method we have successfully generated both over-expression and knock-down mice models. This novel technique of making transgenic animal is simple, fast, cost effective and minimally invasive.}, URL = {https://www.biorxiv.org/content/early/2016/04/18/049239}, eprint = {https://www.biorxiv.org/content/early/2016/04/18/049239.full.pdf}, journal = {bioRxiv} }