CRISPR-based Diagnostics: Gross Errors, Useful Specificity, and Microfluidic Assays

Wednesday, 29 November 2023 at 10:00

Add to Calendar ▼2023-11-28 00:00:002023-11-28 01:00:00Europe/LondonTitle to be Confirmed.Lab-on-a-Chip and Microfluidics World Congress 2023 in Laguna Hills, CaliforniaLaguna Hills,

Molecular diagnostics based on clustered regularly interspaced short palindromic repeats (CRISPR) enzyme systems have been the subject of intense research and development. CRISPR-associated (Cas) enzyme assays are easily reconfigurable to different nucleic acid targets, highly specific, and compatible with simple kits and microfluidic components. We are conducting studies of the basic CRISPR enzyme kinetics.  We discovered that the great majority of all CRISPR enzyme kinetics studies show data that are inconsistent and which grossly violate basic rules of mass conservation and rate laws. This widespread inconsistency makes it difficult to assess the potential of CRISPR as a diagnostics tool. Following up on this, we quantified the kinetics of a range of CRISPR-Cas systems and demonstrated how these kinetics fundamentally limit detection sensitivity. We also performed a study of CRISPR specificity to small mutations, including single-nucleotide polymorphisms. We are currently developing assays that leverage CRISPR specificity for cancer detection. We will also present a review of microfluidic CRISPR assays and report on our CRISPR assays using on-chip electric field control with a method called isotachophoresis (ITP).

Juan Santiago, Charles Lee Powell Foundation Professor, Stanford University

Juan Santiago

Juan G. Santiago received his MS and PhD in Mechanical Engineering from the University of Illinois at Urbana-Champaign in 1995. He holds the Charles Lee Powell Foundation professorship at Stanford and serves as Vice Chair of the Mechanical Engineering Department. His research includes the development of microsystems for on-chip biochemical analysis, microfluidic cell assay systems, two-phase flow devices, and capacitive deionization technologies. Applications of this work include molecular diagnostics, basic studies of cell biology, electronics cooling, and the production of drinking water. He is a Fellow of the American Physical Society, a Fellow of the American Society of Mechanical Engineering, and a Fellow of the American Institute for Medical and Biological Engineering. In 2022, he was elected to the American Academy of Arts and Sciences and to the National Academic of Inventors. He serves as the Editor-in-Chief of the journal Flow. His work is cited about 1500 times per year (Google Scholar h index of 86). He has authored and co-authored over 220 journal papers and 250 conference papers, and he is a named inventor in 58 issued patents (26 of which are currently licensed).