RNA-based regulation of gene expression
RNA molecules that do not encode proteins (ncRNA) are key regulators of cellular responses. The DNA sequence of genes results in multiple mRNA and ncRNA isoforms. The Marquardt lab (PLEN) has mapped this diversity in the plant Arabidopsis by genome-wide transcript isoform mapping techniques. We identified prominent short-promoter proximal RNAs (sppRNAs) at the ARGONAUTE1 (AGO1) gene. AGO1 is the core component of the RNA Induced Silencing Complex that uses microRNAs to repress complementary mRNA, and is a major focus of studies in the Brodersen lab (BIO).
Your project operates at the cross-section between the Brodersen and Marquardt labs. You will test the hypothesis that sppRNA isoforms of the AGO1 gene contribute to regulation of AGO1 expression, for example in response environmental change (i.e. pathogen perception). You will address this question using CRISPR/Cas9-based genome engineering to make targeted sppRNA deficient AGO1 loci, gene expression analyses, molecular cloning, imaging and functional assays of AGO1. We anticipate that your project will have elements performed in both labs, which will give you the opportunity to experience more experimental strategies and research directions.
Peter Brodersen (email@example.com), Sebastian Marquardt (firstname.lastname@example.org).
Methods used:CRISPR/Cas9-based genome engineering, gene expression analyses, molecular cloning, imaging and functional assays of AGO1 (RNA virus tests).
Keywords:RNA, CRISPR/Cas9, gene expression, imaging
Supervisor(s):Peter Brodersen (email@example.com), Sebastian Marquardt (firstname.lastname@example.org).