Piyush JainAssistant Professor
MY RESEARCH GROUP IS GENERATING INSIGHTS AND SOLUTIONS TO problems with genome engineering, specifically CRISPR/Cas systems. Over the past few years, the slow-progressing field of genome engineering has been transformed by the breakthrough of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) with astronomical applications in science, medicine, agriculture, biotechnology, and biomanufacturing. Originally derived from the bacterial immune system, the CRISPR/Cas9 technology works by introducing two components inside cells, a Cas9 nuclease that acts like molecular scissors and a guide RNA (sgRNA) that binds with Cas9 and directs the complex to the target DNA to create double-stranded cuts in the DNA. Due to its ease of use, it is becoming a standard tool for genome engineering and the toolbox is exponentially increasing with other variants of CRISPR/Cas systems with applications in DNA and RNA manipulation. The biggest challenges for CRISPR/Cas technology that are affecting the bridge between in vitro and in vivo applications are safety, efficacy, and delivery. To address these pressing concerns, the Jain lab is focused on developing a multi-scale biomolecular engineering platform using nucleic acids chemistry, protein engineering, and nanoengineering. Specific examples include:
UNDERSTANDING AND IMPROVING SPECIFICITY CRISPR/Cas9 can tolerate several mutations in the DNA resulting into undesirable off-target cleavage. What if we change the length and chemistry of the guide RNA? What if we can control the degradation of the CRISPR/Cas complex immediately after it cuts the on-target DNA? Our primary goal is to understand the molecular basis of this issue to be able to engineer CRISPR/Cas systems with improved specificity by modifying its components using nucleic acids design and protein engineering. We employ an array of bioanalytical techniques with immediate applications for the detection and treatment of genetic disorders.
TARGETED DELIVERY OF CRISPR/CAS SYSTEMS Despite the vast literature highlighting the delivery issues with CRISPR/Cas systems, it remains a major concern. How can we get large molecules like Cas9 protein and sgRNA inside the nucleus of a cell? How can we protect the components from degradation or immune response? The answer lies in developing safe and effective non-viral delivery methods. We aim to design multifunctional targeted nanoparticle systems that can protect CRISPR/Cas from degradation and target specific tissues in vivo with immediate applications for detection and treatment of cancer.
Ph.D., 2013, University of Missouri-Kansas City
B.Pharm., 2006, Dr. Hari Singh Gour University (Central University)
Nucleic acids & Protein Engineering
- P. K. Jain and S. H. Friedman. The ULTIMATE Reagent: A Universal Photocleavable and Clickable Reagent for the Regiospecific and Reversible End Labeling of Any Nucleic Acid. ChemBioChem 2018; 19, 1264.
- Jain PK, Ramanan V, Schepers AG, Dalvie N, Panda A, Fleming HE and Bhatia SN. Development of light-activated CRISPR using photocleavable protectors of guide RNAs. Angewandte Chemie International Edition, 2016; 55, 12440.
- Sarode BR, Jain PK and Friedman SH. Polymerizing Insulin with Photocleavable Linkers to Make Light-Sensitive Macropolymer Depot Materials. Macromolecular Bioscience, 2016; 16 (8): 1138-1146.
- Dudani JS, Jain PK, Kwong GA, Stevens KR and Bhatia SN. Photoactivated Spatiotemporally-Responsive Nanosensors of in Vivo Protease Activity. ACS Nano, 2015; 9(12), 11708-11717.