CRISPR-Cas silencing of Apol11b in Murine Erythroleukemia Cells

Dhruv Prasad and Remeeza Ashrafally

Lab Workbench in Farber Hall, East lab.

Workbench in the East lab of Farber Hall.

Undergraduate Student Project

Introduction

CRISPR Cas 9 is a unique gene engineering technology that enables researchers to edit and splice specific portions of the genome. This project used this technology to study the function of an unknown gene in mouse leukemia cells.

We are Dhruv Prasad and Remeeza Ashrafally, and we are both senior undergraduate students in the Biotechnology program. Under supervision of Dr. Stephen Koury, we conducted research on the CRISPR-Cas9 project in Farber Hall, UB south campus. Our job was to isolate the appropriate plasmids using column chromatography kits and analyze them on agarose gels for further research.

While there has been significant research on erythroid differentiation, the function of the gene Apolipoprotein L11b has largely remained unknown. This gene is present in all mammals, and previous experiments with interfering RNA show that it may play a role in hemoglobin production. By utilizing CRISPR-Cas9 technology, we can more accurately observe the effects that Apol11b has on erythroid differentiation hence it could help in the development of future therapeutics.

Abstract

Apolipoprotein L11b gene is known to be transiently expressed during the process of erythroid terminal differentiation, however it's exact function is unknown. The knockout of Apol11b gene may have an effect in the synthesis of heme and erythrocyte differentiation, which may aid in the development of therapeutics. Murine Erythroleukemia cells that have had their Apol11b gene deleted were anticipated to have less expression of heme, which will be observed using morphological and other methods of analyzing expression. This experiment will allow us to learn more about erythrocyte differentiation and the role the Apol11b gene plays, and could give an insight into the pathophysiology of various psychiatric conditions.

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