RNA-guided genome editing with the CRISPR-Cas9 system has great potential for basic and clinical research, but the determinants of targeting specificity and the extent of off-target cleavage remain insufficiently understood.
Using chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq), we mapped genome-wide binding sites of catalytically inactive Cas9 (dCas9). By using multiple gRNAs with improved specificity, we run the large-scale in-vivo and in-vitro CRISPR knockout screenings in pancreatic ductal adenocarcinoma cells. We identified genes whose genetic deletion or pharmacologic inhibition synergistically increase the cytotoxicity of MEK signaling inhibitors. To eliminate the false-positive results from screening studies, CRISPR-STOP method was developed alternative to wild-type Cas9 for gene knockout studies.
Kuscu earned his bachelors degree in the Department of Molecular Biology and Genetics at Middle East Technical University, Ankara Turkey. He continued his academic career in the same field by receiving his master's of science from Bilkent University, Turkey. After his training in his home country, he moved to the United States where he started a Ph.D. program at the Stony Brook University, NY.
Kuscu unraveled the function of the uncharacterized protein, known as KIAA1199, in his Ph.D. studies. His efforts helped the scientific community to understand the role of this protein in the cell migration and this gene has been known as CEMIP (cell migration inducing protein). He also discovered the epigenetic regulation mechanism of this gene in breast cancer settings, and contributed ten peer-reviewed journals in his graduate years.
Kuscu has continued his academic career as a post-doc research associate at Dr.Adli’s Lab in the University of Virginia. He published his first author paper just after a year in Nature Biotechnology where he discovered the off-targets of Cas9 at genomic level. Recently, he found an alternative way to knock out gene by introducing early stop-codons in the exome by using CRISPR-Cas9 technology. He is using these technologies to study the role of chromatin regulators (CRs) in cancer.
Kuscu is also leading a project to explore the spatial and temporal roles of epigenetic marker for precise gene regulation in mammalian systems. So far, Dr.Kuscu contributed seven papers in his four year post-doc period.
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