6/4/2023 0 Comments Macvector attb recombination![]() ![]() Furthermore, expression of the gene of interest is susceptible to positional effects. However, the copy number and location of a transgene inserted into the host genome typically cannot be controlled because it is a random event. Tg animals have been successfully generated by microinjection of genes of interest into the pronuclei of fertilised eggs, allowing the generation of transgenic animals in many species including mouse, rat, cow and pig. 2014), and can function as animal models of human diseases (Masliah et al. Transgenic (Tg) animals are powerful tools for investigating gene function (Olive et al. Thus, the findings suggest that this new Tc strategy employing mES cells and a MI-MAC vector is efficient and useful for animal transgenesis. As an example for the generation of Tc mice with a desired gene by the MI-MAC mES approach, a Tc mouse strain ubiquitously expressing Emerald luciferase was efficiently established. Chimeric mice with high coat colour chimerism were generated from the MI-MAC mES cell lines and germline transmission from the chimera was observed. The recombination rate of the GFP gene at each attachment site (FRT, PhiC31attP, R4attP, TP901-1attP and Bxb1attP) on MI-MAC was greater than 50 % in MI-MAC mES cells. In this study, we constructed mES cell lines containing a MI-MAC vector to directly insert a gene of interest into the MI-MAC in mES cells via a simple transfection method for Tc mouse generation. However, the efficiency of MMCT from CHO to mES cells is very low (<10 −6). To generate transchromosomic (Tc) mice containing a MI-MAC with genes of interest, the desired genes were inserted into MI-MAC in CHO cells, and then the MI-MAC was transferred to mouse embryonic stem (mES) cells via microcell-mediated chromosome transfer (MMCT). Previously, a MAC containing multi-integration site (MI-MAC) was generated to facilitate transfer of multiple genes into desired cells. The mouse artificial chromosome (MAC) has several advantages as a gene delivery vector, including stable episomal maintenance of the exogenous genetic material and the ability to carry large and/or multiple gene inserts including their regulatory elements. ![]()
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