CORE 2: Genetic Engineering Core

Massachusetts General Hospital, Boston MA

Investigators

Linked publications, trials & patents

Abstract

PROJECT SUMMARY Genome engineering technologies permit the alteration of DNA sequences or changes in gene expression, enabling the creation of modified models to test biological hypotheses in living cells and organisms. The mission of our Genetic Engineering (GE) Core is to design and optimize genome and epigenome editing strategies to help define the relationships between hematopoiesis and the progression of cardiovascular diseases (CVDs), which remain a leading cause of death. Our wider Program Project will bring together field-leading researchers to close this gap in knowledge by elucidating and investigating factors that connect hematopoiesis and CVD. Thus, there is tremendous opportunity to address these outstanding questions by leveraging leading-edge technologies like genome and epigenome editing tools to engineer hematopoietic stem and progenitor cells (HSPCs) in vitro or to create new mouse models to investigate CVD. The GE Core will play a collaborative dual role in this consortium. First, the primary function of our Core is to serve the Projects using pre-existing GE tools by designing, optimizing, and implementing a variety of editing approaches for use in HSPCs (prior to transplant into live recipient mice) or to create novel animal models, both of which will expedite our PPG objectives. We will implement streamlined in silico and experimental methods to design and validate GE strategies to test hypotheses from all Project teams, will optimize gene editing strategies to knockout genes in cells or construct new animal models (Projects 2-4), will deploy epigenetic methods to alter gene expression (Project 1), and will assist with the design and validation of CRISPR screens (Project 4). Our GE Core will implement various GE technologies capable of a diversity of edit types (e.g. nuclease, base editors, prime editing, or sequence insertion methods) to perturb or install candidate genes nominated by Projects 1-4. The second mission of our Core is to develop next-generation technologies that overcome key issues of CRISPR-based GE methods, further enabling our key experimental goals. To do so, we will utilize a suite of protein engineering methods to develop novel CRISPR-Cas editors with improved properties (e.g. enzymes with enhanced on-target efficiency), with expanded scopes of edit outcomes and edit sizes (e.g. large insertions), all while maintaining stringent genome-wide safety in HSPCs and other primary cell types. Our collaborative efforts with the Projects will place an emphasis on innovation to expedite the key objectives of this Program. The Projects will have immediate access to the most advanced state-of-the-art editing technologies and methods, which together will enable rapid, efficient, and safe genetic edits to investigate the connections between hematopoiesis and CVD. More broadly, these advances will be applicable to the study of cardiovascular and hematopoietic disorders, with a path to new therapeutics.