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Identifying novel Cas variants for pathogen diagnostics

Project

Project Details

Program
BioEngineering
Field of Study
Bioengineering
Division
Biological and Environmental Sciences and Engineering

Project Description

Rapid, point-of-care (POC) diagnostics are essential to mitigate the impacts of current (and future) epidemics; however, current methods for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) require complicated laboratory tests that are generally conducted off-site and require substantial time. CRISPR-Cas systems have been harnessed to develop sensitive and specific platforms for nucleic acid detection. These detection platforms take advantage of CRISPR enzymes’ RNA-guided specificity for RNA and DNA targets and collateral trans activities on single-stranded RNA and DNA reporters. Microbial genomes possess an extensive range of CRISPR enzymes with different specificities and levels of collateral activity; identifying new enzymes may improve CRISPR-based diagnostics. We work to identify new Cas variants, and characterize its catalytic activity.

About the Researcher

Magdy Mahfouz
Professor, Bioengineering
Biological and Environmental Science and Engineering Division

Affiliations

Education Profile

  • Ph.D. Molecular Genetics, The Ohio State University, Columbus Ohio, USA 2004
  • M.S. Microbial Genetics, Cairo University, Cairo Egypt, 1997

Research Interests

Professor Mahfouz's research interests are focused on developing genome-engineering technologies for basic biology and biotechnology. The application of genome editing technologies requires highly specific and customizable DNA binding modules that can be engineered to bind any user-defined DNA sequence. Transcriptional activator-like effectors (TALEs) are proteins secreted by Xanthomonas bacteria when they infect plants. TALEs contain a modular DNA binding domain that can be easily engineered to bind any sequence of interest, and have been used, by our group and others, to provide user-selected DNA-binding modules to generate chimeric nucleases and transcriptional regulators in mammalian cells and plants. TALE DNA binding modules fused with endonucleases (TALENs) can direct nuclease activity to site-specific sequences in the genome with extreme precision, allowing targeted gene knock out, integration and correction. Developing TALE-based technologies will allow researchers to routinely and efficiently edit genomes of virtually any species, by directing mutations in a truly targeted fashion.

Selected Publications

  • Ahmed Mahas, Tin Marsic, Mauricio Lopez-Portillo Masson, Qiaochu Wang, Rashid Aman, Cheng Zheng, Zahir Ali, Madain Alsanea, Ahmed Al-Qahtani, Bernard Ghanem, Fatimah Alhamlan, Magdy Mahfouz (2022). Characterization of a thermostable Cas13 enzyme for one-pot detection of SARS-CoV-2. Proceedings of the National Academy of Sciences. 119, 28.
  • Tin Marsic, Zahir Ali, Muhammad Tehseen, Ahmed Mahas, Samir Hamdan, and Magdy M. Mahfouz* (2021). Vigilant: An Engineered VirD2-Cas9 Complex for Lateral Flow Assay-Based Detection of SARS-CoV2. Nano Letters 2021 21 (8), 3596-3603.
  • Zahir Ali, Ashwag Shami, Khalid Sedeek, Radwa Kamel, Abdulrahman Alhabsi, Muhammed Tehseen, Norhan Hassan, Haroon Butt, Ahad Kababji, Samir Hamdan andMagdy M. Mahfouz* (2020). Fusion of the Cas9 endonuclease and the VirD2 relaxes facilitates homology-directed repair for precise genome engineering in rice. Communications Biology 2020 Jan 23; 3(1):44.
  • Ahmed Mahas, Rashid Aman, and Magdy Mahfouz* (2019) CRISPR-Cas13d mediates robust RNA virus interference in plants. Genome Biology, Dec 2; 20(1):263. doi: 10.1186/s13059-019-1881-2
  • Rashid Aman, Zahir Ali, Haroon Butt, Ahmed Mahas, Fatimah Aljedaani, M. Zuhaib Khan, Shouwei Ding, and Magdy M. Mahfouz*(2018) RNA virus interference via CRISPR/Cas13a system in plants. Genome Biology 19: (1). P1-9 (DOI 10.1186/s13059-017-1381-1)

Desired Project Deliverables

Training on different molecular biology techniques and comprehending the components and mechanisms of the CRSIPR-Cas systems.