The effect of suppressing HIF1A on the expression of HIF2A

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The effect of suppressing HIF1A on the expression of HIF2A Project Author: Shray Amin Project Mentor: Keith Baker

Background Information A transcription factor is a protein that binds to specific DNA sequences, thereby controlling the rate of transcription of genetic information from DNA to messenger RNA transcription factors that creates a response to hypoxia

Implications HIFs are known to known to transcribe more than 60 genes, which affect biological processes such as angiogenesis and regeneration of lost or damaged tissue. A better understanding of the interplay between HIF 1 and 2 alpha can offer valuable incite into research in regenerative treatments and cancer growth.

Project Aim To determine the consequential effects on HIF2a via the suppression of HIF1a HIF-2a ?

The Experiment The template for the experiment will follow Rosenberger et al, (2002). Two groups of rat Kidneys kept in low oxygen chambers to induce hypoxia One group will be treated with HAF, which will knockout HIF1a Both groups will under go immunohistochemical analysis to measure the relative expression of the two different HIFs

Hypoxia Associated Factor (HAF) HAF is a protein that ubiquinatesHIF1a, thus marking it for proteasomal destruction. HAF will be incorporated in the form of a plasmid modeled after Koh et al. (2008). pOTB7

Immunohistochemistry the process of detecting antigens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues

Potential Outcome During the first 24 hours that a cell is exposed to a Hypoxic environment, HIF1a is predominantly responsibly for transactivation of target genes. Whereas after 24 hours, HIF2a begins to exert more of an influence (Koh et al., 2011, Koh and Powis, 2012).