Generation of factor VIII gene knockout mouse by tetraploid embryo complementation technology.
- Author:
Ying KUANG
1
;
Jinjin WANG
;
Xibin LU
;
Shunyuan LU
;
Liangliang ZHANG
;
Chunling SHEN
;
Jian FEI
;
Zhugang WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Disease Models, Animal; Embryo, Mammalian; Factor VIII; genetics; metabolism; Female; Hemophilia A; genetics; metabolism; physiopathology; Humans; Male; Mice; Mice, Inbred ICR; Mice, Knockout; Partial Thromboplastin Time
- From: Chinese Journal of Medical Genetics 2010;27(1):1-6
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEFactor VIII( FVIII) gene knockout mouse model was established for further study on the treatment of hemophilia A.
METHODSExons 16-19 of the mouse FVIII gene were knocked out by ET clone, ES homologous recombination and tetraploid embryo compensation technology. PCR, reverse transcriptase-PCR(RT-PCR) and immunohistochemistry were used to detect the transcription and translation pattern of FVIII. The phenotype of the knockout mice was analyzed by examining the activated partial thromboplastin time (APTT) and FVIII activity (FVIII:C).
RESULTSPCR, RT-PCR and immunohistochemistry confirmed that FVIII was deficient in the FVIII gene knockout mouse. The APTT results showed that FVIII-deficient mouse plasma had a prolonged clotting time compared to normal mouse plasma. The FVIII:C in heterozygous, hemizygous and homozygous mice was 80%, 8% and 10% of that in normal mice, respectively.
CONCLUSIONThe phenotype of the FVIII gene knockout mouse appears grossly similar to that of human with hemophilia A. Establishment of this model may promote the development of new technologies of treatment to hemophilia A.
