OVERVIEW

The physiological effects of cancer are a manifestation of the genetic abnormalities that cause the disease. While much progress has been made in the understanding of such genetic perturbations, scientists still struggle to effectively identify, understand, and treat cancer-causing mutations. This is due to the fast-paced evolution of the disease, and the accumulation of novel mutations that permit cell survival even in the harsh environment created by a therapeutic.

CRISPR is a gene-editing technology that couples the elegance of base complementarity with the enzymatic activity of a DNA nuclease in order to introduce mutations into target loci.

CRISPR APPLICATIONS

High-throughput Screening

We discovered that targeting exons encoding critical protein domains results in a more robust target knockdown, and thus enhanced high-throughput screening data. Our approach can help to identify alleles that will confer drug resistance prior clinical use of the drug. This provides an opportunity to establish drug combinations that will prevent resistance, and thus enhance overall treatment efficacy.

In addition to performing CRISPR screens, our lab is interested in investigating and engineering related gene-editing proteins for use in screenings, mutagenesis, and other novel applications.

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FEATURED PROJECTS

NOMINATION OF NOVEL GENETIC DEPENDENCIES IN LEUKEMIA

Our lab has several CRISPR libraries targeting a wide array of basic cell function, including the epigenome and the kinome. In a negative selection screen, we look for crRNAs that exhibit a great dropout (log2 fold change) in their representation over time, suggesting that the genes they target are essential for the cells' survival. By conducting CRISPR screens, we are able to nominate novel genetic dependencies that may provide a therapeutic avenue for the treatment of leukemia. In addition to nominating a genetic vulnerability, we work to uncover the underlying biological mechanism in an effort to further our understanding of leukemia initiation, maintenance, and progression.

ADAPTATION OF ASCAS12A FOR USE IN COMBINATORIAL CRISPR SCREENING

CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. We developed a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we achieved high efficiency combinatorial genetic screening. We successfully demonstrate the performance of our optimized AsCas12a (opAsCas12a) through double knockout screening against epigenetic regulators. From these screens, we were able to identify three potential synthetic sick interactions in a murine leukemia.

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