Tn5 transposase-mediated mouse transgenesis.
We have developed a novel method for mouse transgenesis. The procedure relies on a hyperactive Tn5 transposase to insert a transgene into mouse chromosomes during intracytoplasmic sperm injection. This procedure integrates foreign DNA into the mouse genome with dramatically increased effectiveness as compared to conventional methods such as pronuclear microinjection and traditional sperm injection-mediated transgenesis. Our data indicate that with this method, transgenic mice, both hybrids and inbreds, can be produced more consistently and with lower numbers of manipulated oocytes required for traditional microinjection methods. The transposase-mediated transgenesis technique is also effective with round spermatids, offering the potential for rescuing the fertility of azoospermic animals using sperm precursor cells.
Suganuma
Ryota
R
Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu, Hawaii 96822, USA.
Pelczar
Pawel
P
Spetz
Jean François
JF
Hohn
Barbara
B
Yanagimachi
Ryuzo
R
Moisyadi
Stefan
S
D000818:Animals; D015139:Blotting, Southern; D005260:Female; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D008297:Male; D008540:Meiosis; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic / Q000235:genetics*; D008845:Microinjections; D009865:Oocytes / Q000502:physiology; D016133:Polymerase Chain Reaction; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods; D019076:Transgenes; D019895:Transposases / Q000235:genetics* / Q000378:metabolism
C473503:Tn5 transposase; D019895:Transposases