Generation of transgenic mice and germline transmission of a mammalian artificial chromosome introduced into embryos by pronuclear microinjection.
We have generated transgenic mice by pronuclear microinjection of a murine satellite DNA-based artificial chromosome (SATAC). As 50% of the founder progeny were SATAC-positive, this demonstrates that SATAC transmission through the germline had occurred. FISH analyses of metaphase chromosomes from mitogen-activated peripheral blood lymphocytes from both the founder and progeny revealed that the SATAC was maintained as a discrete chromosome and that it had not integrated into an endogenous chromosome. To our knowledge, this is the first report of the germline transmission of a genetically engineered mammalian artificial chromosome within transgenic animals generated through pronuclear microinjection. We have also shown that murine SATACs can be similarly introduced into bovine embryos. The use of embryo microinjection to generate transgenic mammals carrying genetically engineered chromosomes provides a novel method by which the unique advantages of chromosome-based gene delivery systems can be exploited.
Co
D O
DO
Chromos Molecular Systems, Inc., Burnaby, British Columbia, Canada. dco@chromos.com
Borowski
A H
AH
Leung
J D
JD
van der Kaa
J
J
Hengst
S
S
Platenburg
G J
GJ
Pieper
F R
FR
Perez
C F
CF
Jirik
F R
FR
Drayer
J I
JI
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000235:genetics*; D002875:Chromosomes / Q000235:genetics*; D004276:DNA, Satellite / Q000235:genetics*; D004622:Embryo, Mammalian; D005260:Female; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008845:Microinjections; D009865:Oocytes / Q000166:cytology*; D016133:Polymerase Chain Reaction
D004276:DNA, Satellite