MIT CompBio Lecture 24 - Genome Engineering

MIT Computational Biology: Genomes, Networks, Evolution, Health Prof. Manolis Kellis http://compbio.mit.edu/6.047/ Fall 2018 Lecture 24 - Genome Engineering 1. High-throughput synthesis: Massively Parallel Reporter Assays (MPRA) - MPRA technology: in vitro synthesis, reporter design, transfection - 1 site X deep: single-base dissection, combinatorial changes, high-res map - 2k sites x shallow: motif-guided perturbations, activators/repressors 2. Next-generation applications of MPRA + related technologies - 10k x deep: high-resolution dissection by tiling + deconvolution (Sharpr) - 10M x deep: HiDRA, no synthesis (STARR), targeted capture (ATAC) 3. Endogenous genome editing: cutting and template-based repair - Endonucleases: TALENs, ZNFs. Repair pathways. Diversity of outcomes. - CRISPR-Cas9: origins, discovery, optimization, base-editing - Next-generation: multiplexing, delivery, applications Slides for Lecture 24: https://stellar.mit.edu/S/course/6/fa18/6.047/courseMaterial/topics/topic2/lectureNotes/Lecture25_GenomeEngineering_CRISPRCas9/Lecture24_GenomeEngineering2.pdf