CRISPR/CAS9 TO STUDY PEPTIDE INDUCED HISTONE MODIFICATIONS IN LEGUMES

The overall objective of this project is to understand peptide hormone control of the Medicago truncatula nodule methylome using advanced molecular tools such as gene editing and CHIP seq (chromatin immunoprecipitation (ChIP) with massively parallel DNA sequencing).The project will have the following three objectivesObjective 1: Catalogue changes to the status of the nodule methylome upon peptide treatment.Objective 2: Characterize four core chromatin remodeling enzymes targeted by peptide signaling during nodulation. 2.1 Develop stably transformed overexpression lines and gene-edited lines in M. truncatula for one Histone Methyl Transferase and one Histone Acetyl Transferase. 2.2 Perform RNAseq using two peptides on control and gene edited lines to determine extent of change to the nodule epigenome. 2.3 Identify nodulation-regulated targets of histone modifying enzymes upon peptide treatments usingChromatin Immuno-precipitation (ChIP-seq).Objective 3: Train two graduate students and upto six undergraduate students in advanced biotechnological tools.