Session 3J: Targeting MLL Gene Expression using CRISPR/Cas9 to Reverse the Phenotypes of Cancer

Session Number

Session 3J: 1st Presentation

Advisor(s)

Dr. Vandana Chinwalla, IMSA

Location

Room A123

Start Date

26-4-2018 12:40 PM

End Date

26-4-2018 1:25 PM

Abstract

Mixed Lineage Leukemia (MLL) is a gene that is frequently mutated in many forms of childhood leukemias and solid tumors. MLL is a histone methyltransferase and plays an important role in gene expression of downstream targets by modulating chromatin structure. When MLL is mutated, it results in excessive growth and the formation of cancer. Previous research suggests that at least one normal copy of MLL may be required for the formation of tumors and its aggressive nature through its expression. The project utilizes the gene-editing tool CRISPR/Cas9 tool to knock out the MLL gene in MCF-7 (breast cancer) cells to reverse the phenotypes of cancer, starting apoptosis. A MLL-guideRNA construct was designed for the CRISPR/Cas9 mechanism to target the desired gene. Through transfection, the CRISPR construct with guideRNA, and selection markers were introduced into the cells. Our preliminary results showed that knocking out the MLL gene in MCF-7 cells increased cell death. After attempting to recreate our results, our cells underwent apoptosis earlier than expected. This occurred in multiple trials, thus we aim to discover the underlying cause. The effect of MLL knockout will be evaluated in other tumor lines. The applications of this project can lead to major changes in cancer treatment and further research on MLL’s and cancer.

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Apr 26th, 12:40 PM Apr 26th, 1:25 PM

Session 3J: Targeting MLL Gene Expression using CRISPR/Cas9 to Reverse the Phenotypes of Cancer

Room A123

Mixed Lineage Leukemia (MLL) is a gene that is frequently mutated in many forms of childhood leukemias and solid tumors. MLL is a histone methyltransferase and plays an important role in gene expression of downstream targets by modulating chromatin structure. When MLL is mutated, it results in excessive growth and the formation of cancer. Previous research suggests that at least one normal copy of MLL may be required for the formation of tumors and its aggressive nature through its expression. The project utilizes the gene-editing tool CRISPR/Cas9 tool to knock out the MLL gene in MCF-7 (breast cancer) cells to reverse the phenotypes of cancer, starting apoptosis. A MLL-guideRNA construct was designed for the CRISPR/Cas9 mechanism to target the desired gene. Through transfection, the CRISPR construct with guideRNA, and selection markers were introduced into the cells. Our preliminary results showed that knocking out the MLL gene in MCF-7 cells increased cell death. After attempting to recreate our results, our cells underwent apoptosis earlier than expected. This occurred in multiple trials, thus we aim to discover the underlying cause. The effect of MLL knockout will be evaluated in other tumor lines. The applications of this project can lead to major changes in cancer treatment and further research on MLL’s and cancer.