Molecular Cloning and Functional Characterization of the CBX7 Transcript Variant Gene Open Access

Abstract

Polycomb group proteins (PcG) are epigenetic regulators that modify chromatin structures and control gene expressions. These proteins play critical roles in cancer development. CBX7, a subunit of PcG, has been previously identified to play two opposing roles: a tumor suppressor and an oncogene. Regarding this discrepancy, it was suggested that the function of CBX7 could be tissue- and context-specific. In this study, we identified the Cbx7 variant gene that may possibly explain the discrepancy of functions of CBX7. We successfully cloned Cbx7 variant gene via TA cloning. The sequencing result indicated that Cbx7 and variant gene have different exon 5 region. Based on this data, we hypothesized that the Cbx7 variant gene that contains the extra exon 5 region is the controlling factor in whether the Cbx7 gene will become an oncogene or tumor suppressor. For the gain of function (GOF) analyses, we constructed recombinant adenovirus that overexpresses Cbx7 and its variant gene. These adenoviral particles were used to infect the mouse embryonic fibroblasts (MEF). To validate and characterize GOF effects, we performed quantitative real-time PCR to check expression, Western blot to check the protein expression, and MTT assay to indirectly check cell proliferation rate. Understanding the mechanisms underlying the discrepancy on CBX7 function will grant scientists a clearer understanding of its function and will assist in the development of future studies regarding developmental biology, cancer treatment, tissue homeostasis, and tissue regeneration. 

Table of Contents

Table of Contents

Introduction………………………………………………………………………………….....1-4

Material and Methods ………………………………………………………………………..4-9

Discussion………………………………………………………………………………….....27-29

References………………….………………………………………………………………....30-34

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Genetics Biology, General Biology, Molecular
Keyword	Polycomb-repressive complexes CBX7 Chromobox family proteins
Committee Chair / Thesis Advisor	Young-sup Yoon , Emory University
Committee Members	Kathleen Campbell , Emory University William G. Kelly, Emory University

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