A non-surgical approach for male germ cell mediated gene transmission through transgenesis
| Autor: | Nirmalya Ganguli, Suryaprakash R. Batta, Suveera Dhup, Nilanjana Ganguli, Manoj Vimal, Abul Usmani, Perumal Nagarajan, Subeer S. Majumdar, Hironmoy Sarkar, S. Basu |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Male
Transgene Gene Expression Mice Transgenic Biology Bioinformatics Article Animals Genetically Modified Mice Genes Reporter Testis medicine Animals Transgenes Microinjection Multidisciplinary Electroporation Gene Transfer Techniques Embryo Male pronucleus Embryonic stem cell Cell biology Transgenesis Germ Cells medicine.anatomical_structure Germ cell |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep03430 |
| Popis: | Microinjection of foreign DNA in male pronucleus by in-vitro embryo manipulation is difficult but remains the method of choice for generating transgenic animals. Other procedures, including retroviral and embryonic stem cell mediated transgenesis are equally complicated and have limitations. Although our previously reported technique of testicular transgenesis circumvented several limitations, it involved many steps, including surgery and hemicastration, which carried risk of infection and impotency. We improved this technique further, into a two step non-surgical electroporation procedure, for making transgenic mice. In this approach, transgene was delivered inside both testes by injection and modified parameters of electroporation were used for in-vivo gene integration in germ cells. Using variety of constructs, germ cell integration of the gene and its transmission in progeny was confirmed by PCR, slot blot and immunohistochemical analysis. This improved technique is efficient, requires substantially less time and can be easily adopted by various biomedical researchers. |
| Databáze: | OpenAIRE |
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