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Cytoplasmic injection of murine zygotes with Sleeping Beauty transposon plasmids and minicircles results in the efficient generation of germline transgenic mice by Wiebke Garrels; Thirumala R. Talluri; Maren Ziegler; Ilka Most; Diego O. Forcato; Marco Schmeer; Martin Schleef; Zoltán Ivics; Wilfried A. Kues is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
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## Abstract Transgenesis in the mouse is an essential tool for the understanding of gene function and genome organization. Here, we describe a simplified microinjection protocol for efficient germline transgenesis and sustained transgene expression in the mouse model employing binary Sleeping Beauty transposon constructs of different topology. The protocol is based on co‐injection of supercoiled plasmids or minicircles, encoding the Sleeping Beauty transposase and a transposon construct, into the cytoplasm of murine zygotes. Importantly, this simplified injection avoids the mechanical penetration of the vulnerable pronuclear membrane, resulting in higher survival rates of treated embryos and a more rapid pace of injections. Upon translation of the transposase, transposase‐catalyzed transposition into the genome results in stable transgenic animals carrying monomeric transgenes. In summary, cytoplasmic injection of binary transposon constructs is a feasible, plasmid‐based, and simplified microinjection method to generate genetically modified mice. The modular design of the components allows the multiplexing of different transposons, and the generation of multi‐transposon transgenic
Who reads Cytoplasmic injection of murine zygotes with Sleeping Beauty transposon plasmids and minicircles results in the efficient generation of germline transgenic mice?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Wiebke Garrels; Thirumala R. Talluri; Maren Ziegler; Ilka Most; Diego O. Forcato; Marco Schmeer; Martin Schleef; Zoltán Ivics; Wilfried A. Kues
- Publisher
- Wiley
- Published
- 2015
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)