Ben Kleinstiver, PhD
Ben Kleinstiver is a biochemist and genome editor whose interests include translating technologies into molecular medicines. He received his Ph.D in Biochemistry from the University of Western Ontario, and completed his postdoctoral studies at Massachusetts General Hospital and Harvard Medical School. Within the Center for Genomic Medicine at MGH, the Kleinstiver laboratory seeks to address limitations of CRISPR genome editing technologies while solving important research questions at the forefront of the genome editing field.
Questions being addressed in the lab
Projects underway to answer these questions
Improving the targeting range and activities of CRISPR genome editing technologies. A few major limitations of genome editing enzymes include: 1) the inability to target sequences of interest, or 2) suboptimal/inadequate activity. We have engineered naturally occurring CRISPR nucleases to enable them to target more frequently and to access previously untargetable sequences in the genome. These engineered enzymes have implications for various genome editing applications including gene knock-out and knock-in, epigenome editing, base editing, and allele specific discrimination.
Defining and minimizing the off-target effects of CRISPR nucleases. Genome editing technologies can exhibit the propensity to target DNA sequences that resemble the intended site. The recognition and cleavage of ‘off-target’ sites can lead to undesirable consequences for research applications, and can have major implications for the therapeutic use of genome editing technologies. To prevent off-target cutting, we have ongoing efforts to develop nucleases with vastly improved genome-wide specificities.
Characterizing and developing alternative technologies with enhanced properties. Beyond the prototypical commonly used SpCas9 nuclease, there is a tremendous variety of enzymes that have distinctive properties. The diverse characteristics of these alternative platforms may therefore be advantageous for biomedical research or potentially for the treatment of disease.
Evolution of CRISPR nucleases with expanded targeting range and enhanced on-target activities.
Manuscripts: https://www.ncbi.nlm.nih.gov/pubmed/26098369 and https://www.ncbi.nlm.nih.gov/pubmed/26524662
Development of safer CRISPR genome editing technologies via protein engineering to reduce or eliminate off-target effects.
Manuscripts: https://www.ncbi.nlm.nih.gov/pubmed/26735016 and https://www.ncbi.nlm.nih.gov/pubmed/28931002
Leveraging technologies in the genome editing ‘toolbox’ with desirable properties to target and correct pathogenic sequences.
Highlighted Publications from the lab
Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing