Submission Title

Investigating a Viral Response Gene Using the CRISPR Editing Process

Location

Jereld R. Nicholson Library: Grand Avenue

Subject Area

Biochemistry

Description

Of many genes that respond to viral infections, most are not well understood. HP, a gene normally involved in hemoglobin recycling, is known to be upregulated after viral infection. In an effort to mutate this gene of interest and better understand the function of this gene, the CRISPR-Cas9 system was used. CRISPR is a gene editing tool that cleaves the targeted DNA of the gene, potentially causing a mutation. A mutation or knockout of the gene could illustrate how the immune response functions in the absence of this upregulated gene. This process was done with two guides (sgRNA) to increase the chances of knocking out the gene and because we had reason to believe both were equally useful. We are certain that the plasmids contain our guides due to results from gel electrophoresis and confirmation by PCR sequencing; we also found that the cells express the plasmid because the cells survived antibiotic selection processes. Primers were designed for both PCR genomic sequencing and confirmation of the mutation. In the future, sequencing and analysis of the DNA and proteins can be done to confirm mutation of HP.

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Investigating a Viral Response Gene Using the CRISPR Editing Process

Jereld R. Nicholson Library: Grand Avenue

Of many genes that respond to viral infections, most are not well understood. HP, a gene normally involved in hemoglobin recycling, is known to be upregulated after viral infection. In an effort to mutate this gene of interest and better understand the function of this gene, the CRISPR-Cas9 system was used. CRISPR is a gene editing tool that cleaves the targeted DNA of the gene, potentially causing a mutation. A mutation or knockout of the gene could illustrate how the immune response functions in the absence of this upregulated gene. This process was done with two guides (sgRNA) to increase the chances of knocking out the gene and because we had reason to believe both were equally useful. We are certain that the plasmids contain our guides due to results from gel electrophoresis and confirmation by PCR sequencing; we also found that the cells express the plasmid because the cells survived antibiotic selection processes. Primers were designed for both PCR genomic sequencing and confirmation of the mutation. In the future, sequencing and analysis of the DNA and proteins can be done to confirm mutation of HP.