School of Engineering Department of Chemical and Biological Engineering 78 Department of Chemical and Biological Engineering CRISPR-Cas Systems for Disease Diagnostics and Therapeutics Supervisor: HSING I-ming / CBE Student: CAO Jinhui / BIEN Course: UROP 1100, Summer Discovered as a part of the immune system in bacteria, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and its associated proteins (Cas proteins) have been widely studied and engineered, demonstrating great capability for nucleic acid detection and cleavage. While the CRISPR/Cas systems that exist in nature and are studied in most research use RNA-guided DNA-targeting machinery, our previous study overturned this paradigm with a DNA-guided RNA-targeting platform. This report will focus on the UROP project that attempt to extend the DNA-guided methodology to other CRISPR/Cas systems by examining binding affinity and cleavage ability. Optimization on the systems’ reaction rate through additives screening will be demonstrated. It will also discuss the exploratory process in research. Engineering an Orthogonal Recombinant Glycoconjugate Vaccine for Protection Against Enterotoxigenic Toxins Generated by Escherichia Coli Supervisor: HSING I-ming / CBE Student: LI Jizheng / BCB Course: UROP 1100, Summer Shiga toxins, produced by Enterotoxigenic Escherichia coli, are major virulence factors responsible for severe human diseases. This project aims to employ synthetic biology techniques to engineer an orthogonally glycoconjugate vaccine capable of enhancing the neutralizing effect of an antibody against the lethal enterotoxin. Engineering an Orthogonal Recombinant Glycoconjugate Vaccine for Protection Against Enterotoxigenic Toxins Generated by Escherichia Coli Supervisor: HSING I-ming / CBE Student: TANATA Nico / BIOT-AB Course: UROP 1000, Summer One of the causes of food poisoning is the shiga toxin, released from a kind of E.coli known as ‘Shiga Toxinproducing E.coli’ (STEC). By attacking microvilli in our intestines, absorption is affected, and dehydration occurs. The toxin also increases the risk of hemolytic uremic symptom (HUS), leading to renal failure. These effects can prove fatal to infants and the elderly, not to mention that their immune system is either immature or aged, which is why a glycoconjugate vaccine would be more suitable against STEC rather than a regular vaccine, as it will elicit a stronger immune response with the additional peptide molecule. This report will be focusing mainly on the protein purification part in researching a glycoconjugate vaccine against the shiga toxin.
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