The Role of Transcription Factor TFII-I/Gtf2i in Erythro-Megakaryocytic Cell Differentiation

Ryan Fishman and Alexis Trumbull

Authors:  Ryan Fishman, Alexis Trumbull

Faculty Mentor: Jorg Bungert

College:  College of Medicine

Abstract

TFII-I is a transcription factor known to both positively and negatively impact gene expression in cells in response to stress, cell cycle, and cell differentiation. This study analyzes TFII-I function throughout hematopoiesis using mice in which exon 3 of the TFII-I gene was conditionally deleted via the Cre-Lox Recombinase (LoxP) system. We analyzed gene expression and hematopoietic cell counts in the TFII-I depleted and in control mice. Conditional deletion of TFII-I has been associated with a statistically significant decrease in the number of megakaryocytes and has been seen to have a mild effect on red blood cell production. We observed a 2-fold increase in the number of megakaryocytes (n=3). We also generated a DNA construct for the introduction of a biotinylatable tag into the TFII-I coding region in HUDEP2 cells using CRISPR/Cas9 technology. We will use streptavidin pull-down to analyze proteins that TFII-I interacts with or genomic DNA loci associated with TFII-I. TFII-I has been implicated in the formation of lymphomas and our studies may shed light on the function of TFII-I that may be relevant to its role in cancer.

Poster Pitch

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Poster

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16 Responses
    1. Alexis Trumbull

      Thank you Alexis Brake! Next we hope to introduce the constructed biotinylatable tag into the genome of our HUDEP2 cells. Past findings revealed that the depletion of TFII-I was accompanied with an increase in beta-globin gene expression and no change in alpha-globin gene expression. We are hoping that with this new, complete tagging/pull-down process, we will get more defined results. We are very excited to see where this goes!

  1. Dr. Donnelly

    Nice poster layout. When presenting to a general audience it is important to tell them up front why this is an important area to be researching. A professional audience would know but others might not. Impressive work.

    1. Alexis Trumbull

      Thank you for your advice Dr. Donnelly! We are so grateful to be able to use today to learn and better our skills!

  2. Alexis Trumbull

    Thank you Alexis Brake! We next plan on introducing the constructed biotinylatable tag to the genome of the HUDEP2 cells. The technique we have designed will allow us to have a more accurate and complete view of what truly occurs once the gene is pulled down. Prior findings convey that the partial-depletion of TFII-I in these cells leads to a significant increase in beta-globin gene expression while alpha-globin gene expression is unaffected. These more complete findings and analysis of TFII-I function could later be applied to the treatment of several common blood diseases around the world! We are very excited to see where this all goes!

  3. Aishwarya Gurumurthy

    Nice explanation and very well made poster! I see that you have been answering questions well, it’s impressive.

  4. Alicia Hou

    Very interesting work, great job!
    Could just be me, but unfortunately I can’t read some of the smaller print on the poster (I think maybe the uploaded image quality is a little low). Thanks for having explanations to go with each part of the figures, although it’s a bummer I can’t see all the axes and things.