[Establishment of a mutant Lumican transgenic mouse model].
OBJECTIVE
Pathological myopia (PM) is a hereditary ocular disease leading to severe loss of visual acuity and blindness. Lumican gene (LUM) is one of those candidate genes of PM. The purpose of this study was to establish a mutant Lumican transgenic mouse model, and to prepare for the further study of the pathogenesis of PM.


METHODS
Experimental study. Mutation of LUM gene was created by site-directed mutagenesis. Recombinant DNA techniques were used for the construction of the pRP. EX3d-EF1A>LUM/flag>IRES/hrGFP transgene. The gene fragments were microinjected into the zygote male pronuclei of BDF1 mice, and then the zygote cells alive were transplanted into the oviduct of acceptor pregnant female ICR mice. The F0 generation transgenic mice obtained were named C57-TgN (LUM)CCMU. Genome DNA from mice tail was detected by PCR and Western blotting.


RESULTS
Six of 31 F0 generation mice were positive transgenic mice. The western blotting study showed that the flag-tag was expressed in the mouse tail tissue. Sixty-eight of 128 mice (F1 to F3 generation) were positive transgenic mice, the positive rate is 53.13%.


CONCLUSIONS
The mutant Lumican (cDNA 596T>C) transgenic mouse model has been established. This model will provide fundamental conditions for studies of the pathogenesis of PM. Also it will be the basis of further studies about the effect of Lumican mutation on the development of PM and structure and function of the extra cellular matrix.
Beijing Tongren Eye Center,Beijing Tongren Hospital, Capital Medical University, Beijing Key Lab of Ophthalmology & Visual Sciences, Beijing 100730, China.
Yanzheng
Y
Song
Yanyan
Y
Zhao
Email: wxw93@hotmail.com.
Fengju
F
Zhang
Yanqiu
Y
Yu
Ling
L
Ma
D000818:Animals; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics*; D004195:Disease Models, Animal; D005091:Exons; D005260:Female; D007632:Keratan Sulfate / Q000235:genetics*; D000071234:Lumican; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D009216:Myopia / Q000235:genetics / Q000473:pathology*; D010957:Plasmids
D011508:Chondroitin Sulfate Proteoglycans; C000604307:Lum protein, mouse; D000071234:Lumican; D007632:Keratan Sulfate