Apr 18, 2016

Robust generation of transgenic mice by hypotonic shock mediated transgene delivery in testicular germ cells

BioRxiv : the Preprint Server for Biology
Abul UsmaniSubeer Majumdar

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 minimu...Continue Reading

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Mentioned in this Paper

Embryo
Microinjections
Benign Neoplasm of Testis
Research
Genome
Genes
Entire Embryo
Neoplasm of Uncertain or Unknown Behavior of Testis
Tromethamine
Nucleic Acid Sequencing

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