[p16INK4a exon 1 alpha knockout in mouse embryonic stem cells].
INK4a/ARF locus distinguishes itself by its unusual structure and function. It contains 2 overlapping genes with exons 1 alpha, 2 and 3 encoding p16INK4a and exons 1 beta, 2 and 3 encoding p19ARF. Mice with their exons 2 and 3 of the INK4a/ARF knocked out are viable and fertile but develop spontaneous tumors at an early age and highly sensitive to carcinogenic treatment. However, mice with their exon 1 beta knocked out, without interference the expression of p16INK4a, show almost the same phenotype as those with their exons 2 and 3 knocked out. This raises a question of whether the mouse p16INK4a plays a role in tumor suppression. To investigate this problem, a targeting vector pointing to p16INK4a exon 1 alpha with 1.5 kb Eco81 I/Acc II fragment as short arm and 5.9 kb Xba I/Xho I fragment as long arm was built. After linearlization and purification, the targeting vector was introduced into ES cells through electroporation. Thirty-seven G418- and gancyclovir-resistant colonies were picked out and one of them was confirmed as positive by Southern hybridization.
Gong
Zhen-Ming
ZM
Medical Genetics Department, Second Military Medical University, Shanghai 200433, China.
Li
Jian
J
Fu
Ji-Liang
JL
D000818:Animals; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000172:deficiency / Q000235:genetics*; D018274:Electroporation; D004622:Embryo, Mammalian / Q000166:cytology; D005091:Exons / Q000235:genetics; D018390:Gene Targeting; D005822:Genetic Vectors; D051379:Mice / Q000196:embryology; D018345:Mice, Knockout / Q000235:genetics*; D013234:Stem Cells / Q000502:physiology*
D019941:Cyclin-Dependent Kinase Inhibitor p16