BBS Faculty Member - Keith Joung

Keith Joung

Department of Pathology

Massachusetts General Hospital East
Molecular Pathology Unit
149 13th St., 6th Floor
Charlestown, MA 02129
Tel: 617-726-9462
Fax: 617-726-5684
Lab Members: 3 postdoctoral fellows, 3 graduate students, 4 technicians
Visit my lab page here.

The Joung Laboratory develops technologies for genome and epigenome engineering of living cells and organisms using engineered zinc finger, transcription activator-like effector (TALE), and RNA-guided CRISPR-Cas9-based systems and explores their applications for biological research and gene therapy.

Genome Editing Using Targeted Nucleases

Genome editing technology was recently named runner-up for “Breakthrough of the Year” for 2012 and 2013 by Science magazine and “Method of the Year” for 2011 by Nature Methods. We have previously invented two rapid, robust, and publicly available methods for engineering ZFNs known as OPEN (Oligomerized Pool Engineering; Maeder et al., Mol Cell 2008) and CoDA (Context-Dependent Assembly; Sander et al., Nat Methods 2011). In addition, we have also developed and optimized methods for engineering TALENs including an automated, high-throughput method known as FLASH (Fast Ligation-based Automated Solid-phase High-throughput) assembly (Reyon et al., Nat Biotechnol. 2012). We have also recently described reagents that enable the rapid construction of CRISPR-Cas9 nucleases (Hwang et al., Nat Biotechnol. 2013).

Much of our recent work with genome-editing nucleases has focused on CRISPR-Cas9. We and our collaborators were the first to demonstrate that these nucleases can function
in vivo (Hwang & Fu et al., Nat Biotechnol. 2013), modifying endogenous genes in zebrafish and the first to show that they can induce significant off-target mutations in human cells (Fu et al., Nat Biotechnol. 2013). To improve the specificities of these nucleases, we have developed two platforms that show greatly reduced off-target effects: one based on the use of truncated guide RNAs (Fu & Sander et al., Nat Biotechnol. 2014) and the other in which we engineered dimerization-dependent CRISPR-Cas9 nucleases (Tsai et al., Nat Biotechnol. 2014). We recently developed GUIDE-seq, an unbiased, genome-wide method for sensitive detection of CRISPR-Cas9-induced off-target mutations (Tsai et al., Nat Biotechnol. 2015). We have also evolved Cas9 variants with novel DNA binding specificities, thereby broadening the targeting range and applications of this platform (Kleinstiver et al., Nature 2015).

Epigenome Editing Using Targeted Transcription Factors and Epigenetic Modifiers

We have recently demonstrated that the TALE and CRISPR RGN platforms can also be utilized to create artificial customizable transcription factors that can robustly alter expression of endogenous human genes (Maeder et al., Nat Methods 2013a; Maeder et al., Nat Methods 2013b). In addition, we have collaborated with the group of Brad Bernstein to develop fusions of the histone demethylase LSD1 with TALE domains that can induce targeted histone alterations at endogenous human enhancers (Mendenhall et al., Nat Biotechnol. 2013). Finally, we have also developed fusions of engineered TALE domains with the catalytic domain of the TET1 enzyme, enabling the targeted demethylation of CpGs in human cells (Maeder et al., Nat Biotechnol. 2013). We are exploring the use of these and other proteins in both a directed fashion as well as with combinatorial libraries to induce specific phenotypes and cellular states in human cells.

Last Update: 8/10/2015


For a complete listing of publications click here.



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