Efficient metaphase II transgenesis with different transgene archetypes.
Mammalian genome characterization and biotechnology each require the mobilization of large DNA segments to produce transgenic animals. We recently showed that mouse metaphase II (mII) oocytes could efficiently promote transgenesis (mII transgenesis) when coinjected with sperm and small (<5 kilobases) ubiquitously expressed transgenes (tgs). We have extended this work and now report that mII transgenesis can readily be applied to a range of larger tgs (11.9-170 kilobases), including bacterial and mammalian artificial chromosome (BAC and MAC) constructs. The efficiency of large-construct mII transgenesis was at least as high as that with small constructs; 11-47% of offspring carried the large tgs. More than 95% of these transgenic founders transmitted the tg to offspring. These data demonstrate the ability of mII transgenesis to deliver large tgs efficiently.
Perry
A C
AC
The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. tperry@advancedcell.com
Rothman
A
A
de las Heras
J I
JI
Feinstein
P
P
Mombaerts
P
P
Cooke
H J
HJ
Wakayama
T
T
D000818:Animals; D002478:Cells, Cultured; D022202:Chromosomes, Artificial, Bacterial; D022221:Chromosomes, Artificial, Mammalian; D005314:Embryonic and Fetal Development; D005307:Fertilization in Vitro; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008677:Metaphase; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D008845:Microinjections / Q000379:methods; D009865:Oocytes / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology; D019076:Transgenes