Heterochromatin protein 1 deleted chromo domain decreases gene silencing of transgene in mouse.
The heterochromatin protein 1 (HP1) regulates epigenetic gene silencing by promoting and maintaining chromatin condensation. To decrease gene silencing, the chromo domain (CD) in the M31 (the main HP1 in mouse) was deleted by site-directed mutagenesis. Vector pcDNA3.1(+)/M31-DeltaCD, in which the M31-DeltaCD is driven by the CMV promoter, and vector pcDNA3.1(+)/P1A3-M31-DeltaCD, in which the M31-DeltaCD is driven by a goat ss-casein promoter were constructed. The former vector was transfected into a murine fibroblast cell line, which can express enhanced green fluorescent protein (EGFP). EGFP expression, which was determined by flow cytometric analysis, increased approximately 80% in the transfected cells. After injection of the latter vector into transgenic mouse mammary glands, which can express human clotting factor IX (hFIX), the hFIX expression level in the mouse milk increased approximately 40-60% and hFIX in one mouse milk was maintained at a high concentration for over 10 days.
Liu
Feng-Tao
FT
Institute of Medical Genetics, Shanghai Jiao Tong University, 24/1400 West Beijing Road, 200040, Shanghai, PR China. 6ft@sohu.com
Zhang
Yan
Y
D000818:Animals; D019149:Bioreactors; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D005164:Factor IX / Q000096:biosynthesis*; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000096:biosynthesis; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008321:Mammary Glands, Animal / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008892:Milk; D016297:Mutagenesis, Site-Directed / Q000379:methods; D041681:NIH 3T3 Cells; D019076:Transgenes / Q000235:genetics
D002868:Chromosomal Proteins, Non-Histone; D000090266:Chromobox Protein Homolog 5; D049452:Green Fluorescent Proteins; D005164:Factor IX