TY - JOUR
T1 - PITT
T2 - Pronuclear injection-based targeted transgenesis, a reliable transgene expression method in mice
AU - Ohtsuka, Masato
AU - Miura, Hiromi
AU - Sato, Masahiro
AU - Kimura, Minoru
AU - Inoko, Hidetoshi
AU - Gurumurthy, Channabasavaiah B.
PY - 2012
Y1 - 2012
N2 - Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specifc recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-fnger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.
AB - Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specifc recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-fnger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.
KW - Mouse
KW - PITT
KW - Pronuclear injection
KW - Targeted transgenesis
KW - Transgene expression
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UR - http://www.scopus.com/inward/citedby.url?scp=84874049660&partnerID=8YFLogxK
U2 - 10.1538/expanim.61.489
DO - 10.1538/expanim.61.489
M3 - Review article
C2 - 23095812
AN - SCOPUS:84874049660
SN - 1341-1357
VL - 61
SP - 489
EP - 502
JO - Experimental Animals
JF - Experimental Animals
IS - 5
ER -