HR gene silencingBY THE HAIRY BOYS

Introduction

Exposing the scalp to UV light increases risk of skin cancer.

This gene encodes a protein that is involved in hair growth. This protein functions as a transcriptional corepressor of multiple nuclear receptors, including thyroid hormone receptor, the retinoic acid receptor-related orphan receptors and the vitamin D receptors, and it interacts with histone deacetylases. The translation of this protein is modulated by a regulatory open reading frame (ORF) that exists upstream of the primary ORF. Mutations in this upstream ORF cause Marie Unna hereditary hypotrichosis (MUHH), an autosomal dominant form of genetic hair loss. Mutations in this gene also cause autosomal recessive congenital alopecia and atrichia with papular lesions, other diseases resulting in hair loss. Two transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Oct 2014]

hair growth associated transcript variant 1 from Homo sapiens nonfunctional hair growth associated transcript (HR) gene, HR-p.Gln908Ter allele, complete sequence (genbannk: KJ994533.1)

Procedures

1. Choose gene of interest (3.719KBP)

2. PCR – to get ample cDNA count

3. Choose viral vector and RE - pLNCX2 Retroviral Vector EcoRI and cut sites EcoRI & XbaI producing 1.6 kb, 1.9 kb, & 2.6 kb.

4. DNA interest insert to Retrovirus (through transformation)

5. Infect mice with the Retrovirus

6. Allow time to express the gene

1. Mice not bald that means virus does not take up the gene

2. Mice that are hairless virus successfully accepted the gene

7. Hairless mice are then breed to undergo silencing testing

8. SiRNA silencing design – put cDNA into the darmacon

9. Expected results: Hair grows back

Dermacon

Start positon 1105 and 1113

Choosed 80 score and above

Conclusion

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Refernece

http://www.ratbehavior.org/hairless.htm