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Recent progress in biotechnology-based gene manipulating systems to produce knock-in/out mouse modelsopen access

Authors
Lee W.K.Park J.J.Cha S.H.Yun C.-H.
Issue Date
2008
Publisher
Asian-Australasian Association of Animal Production Societies
Keywords
ES cells; Gene manipulation; Knock in/out; ROSA26; Tetraploid; Transgenic
Citation
Asian-Australasian Journal of Animal Sciences, v.21, no.5, pp.745 - 753
Indexed
SCOPUS
KCI
OTHER
Journal Title
Asian-Australasian Journal of Animal Sciences
Volume
21
Number
5
Start Page
745
End Page
753
URI
https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/17385
DOI
10.5713/ajas.2008.r.04
ISSN
1011-2367
Abstract
Gene-manipulated mice were discovered for the first time about a quarter century ago. Since then, numerous sophisticated technologies have been developed and applied to answer key questions about the fundamental roles of the genes of interest. Functional genomics can be characterized into gain-of-function and loss-of-function, which are called transgenic and knock-out studies, respectively. To make transgenic mice, the most widely used technique is the microinjection of transgene-containing vectors into the embryonic pronucleus. However, there are critical drawbacks: namely position effects, integration of unknown copies of a foreign gene, and instability of the foreign DNA within the host genome. To overcome these problems, the ROSA26 locus was used for the knock-in site of a transgene. Usage of this locus is discussed for the gain of function study as well as for several brilliant approaches such as conditional/inducible transgenic system, reproducible/inducible knockdown system, specific cell ablation by Cre-mediated expression of DTA, Cre-ER™ mice as a useful tool for temporal gene regulation, MORE mice as a germ line delete and site specific recombinase system. Techniques to make null mutant mice include complicated steps: vector design and construction, colony selection of embryonic stem (ES) cells, production of chimera mice, confirmation of germ line transmission, and so forth. It is tedious and labor intensive work and difficult to approach. Thus, it is not readily accessible by most researchers. In order to overcome such limitations, technical breakthroughs such as reporter knock-in and gene knock-out system, production of homozygous mutant ES cells from a single targeting vector, and production of mutant mice from tetraploid embryos are developed. With these upcoming progresses, it is important to consider how we could develop these systems further and expand to other animal models such as pigs and monkeys that have more physiological similarities to humans.
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