May 7, 2016, Creative Animodel tells you the easy but useful technology to prepare transgenic animals by electric shock. Rat is the first choice as an animal model, because there are 5 million years of evolution differences between rats and mice. Rats are closer to humans in the genome and physiology. Although the rat is very important in the study of biological medicine, but the production of transgenic animals are mainly using mouse embryos.
Limitations of embryo manipulation
There are many limitations in the production of transgenic rats by embryonic manipulation. Unlike mice, most of the rat embryos could not survive after the injection of a fertilized egg, which reduces the efficiency of producing transgenic rats by this method. More importantly, embryo manipulation requires hundreds of eggs to be inserted to produce transgenic mice, which needs to kill a lot of female rats to get.
By transposon mediated single copy gene insertion, it can improve the efficiency of the standard male pronucleus DNA microinjection, but this method has its own limitations, including the phenomenon of excess suppression. And the operation of the embryo is relatively difficult. Relative to embryo manipulation, embryonic stem cell mediated transgene is another option, but until now the isolation of embryonic stem cells from the rat has been shown to be in vain.
Testis electric shock for transgene
Researchers at the National Institute of immunology in India have developed a non-invasive technique to produce transgenic rats. DNA was injected into the testis to shock. By using no-differentiated rat spermatogonial cells, transgenes would be integrated into the genome of spermatogonial cells by electroporation method. This new technology can obtain transgenic rats through the operation of rat testicular.
By using EGFP (enhanced green fluorescent protein) as the transgene to operate, through the over expression of the beta chain of Hb, transgenic rat disease model successfully shows the similar human alpha – thalassemia. The use of this technology for transgenes and stable integration is in order to deliver to the next generation so that they have a clear pathological phenotype.
Previous studies have reported to prepare gene knockout rats by using CRISPR-CAS9 to transfect male cell line, and the shock of rat testis transformation method for exogenous gene has a high efficiency. The method can directly introduce the gRNA and cas9 RNA directly into the rat spermatogonial cells to produce gene knockout rats. This method is convenient and cost-effective, and does not involve germ cell transplantation.
This study confirmed the feasibility of producing transgenic mice by electrical shock. This effective method will not kill animals and easily produce transgenic rats, and can reduce the number of rats to produce transgenic animals. This simple, less time-consuming and less complex method will help to produce a large number of related animal models in biomedical research.
About the author:
Creative Animodel is specialized in the transgenic animal models for many years and have dedicated to providing the customer with the best services. With the high success rate in generating transgenic services, we have created more than 10.000 animal model with these years.