TATAA, Celiac! A Proposed Model of Celiac DNA Isolation, Intervention and Cure

Voiceover

Presenter(s)(s)

Chloe Pennington

Abstract or Description

• Using a theoretical CRISPRi model is it possible to cancel out the TATAA box code that determines Celiac by causing the mRNA polymerase to scan over it? 
• Independent variable is CRISPR method (CRISPRi or CRISPRko)
• Dependent variable is efficacy of intervention determined by CRISPick 
• My hypothesis is that test one (with HLA-DQ2 and CRISPRi) will be more effective than any other tests; test two (HLA-DQ2 and CRISPR), test three (HLA-DQ8 and CRISPRi), or test four (HLA-DQ8 and CRISPR). I believe that because HLA-DQ2 has the most direct influence over pronouncing more severe Celiac symptoms. HLA-DQ2 is also the more common Celiac gene (it's seen in 90% of people with Celiac Disease), and therefore seen in patients more commonly that are experiencing the aforementioned symptoms. CRISPRi is a novel version of CRISPR that uses genetic perturbation to repress gene expression and therefore the symptoms associated with it. I believe that combining CRISPRi and HLA-DQ2 will create the strongest and most promising method of removal. I hope to discover possible ways for Celiac treatment through a novel gene-editing solution. 
• My prediction is that by changing the promoter sequence of the exon, I will be able to pause the production of the Celiac inducing gene sequence.
• Constants: gene (HLA-DQ8 and HLA-DQ2), editing software (CRISPick), exon and promoter, enzyme (Spyo-Cas9), loci on the chromosome (chromosome 6).
• As you can see from my results, the CRISPRi and HLA-DQ2 had the highest efficacy score of .7397 compared to CRISPRko and HLA-DQ2 (.5911), CRISPRi and HLA-DQ8 (.6374) and CRISPRko and HLA-DQ8 (.6351). My hypothesis was correct, the sgRNA sequence extracted from the HLA-DQ2 combination with CRISPRi creates the most effective method of removal. 
• Methods: I used computer model software (CRISPick) to analyze DNA and protein sequences to understand the influence CRISPR has over the HLA-DQ2 and HLA-DQ8 gene when inserted properly.
• Materials: computer with CRISPick website, isolated gene sequence (found at HLA-DQ2 Marked Up Sequence) 1, and spreadsheet.
• My discoveries help people because of the critical issue Celiac presents. Celiac is an extremely important and relevant issue since many of the Celiac population experience debilitating symptoms. By finding cures to Celiac diseases, we can help the millions who are affected by Celiac disease.