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Regan Miller

Regan Miller
CRISPR-Cas9- mediated gene knockout of defense response receptor in nitrogen-fixing legume model plant
Program Year: 
Biochemistry & Molecular Biology
Dong Wang

This summer I worked in the Wang lab with a focus on learning how to successfully transform plant DNA with CRISPR- Cas9 machinery. The Wang lab studies the symbiotic relationship between legumes and nitrogen-fixing bacteria where the legumes allow the bacteria to infect their roots to form nodules to symbiotically perform nitrogen-fixing. Our lab uses the model legume, Medicago truncatula, and rhizobia to study this molecular process.
I focused on the gene responsible for the LYM2 protein, which is thought to be a defense response receptor that is activated during the infection process. I used CRISPR- Cas9 to target this LYM2 gene and cause a deletion in the gene’s sequence. The first CRISPR vector I designed was unsuccessful in causing a deletion in the LYM2 gene, and the second CRISPR vector was successful in causing the expected deletion. Further experiments can be performed to observe the phenotype of the model legume after the gene’s function has been knocked out due to the CRISPR machinery.