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E-Book 2nd Congress

  • Diagnostic approaches for infectious disease using CRISPR-Cas technology: a review study
  • Zahra Momtahan,1,* Tarlan Arjmandi,2 Fatemeh Ghorbanzadeh,3
    1. Department of Genetics, Faculty of New Sciences and Technologies, Tehran Medical Sciences, Islamic Azad Univeristy, Tehran, Iran
    2. MacCORMIC school of engineering, Master of Science in Biotechnology, Northwestern University, Evanston, Illi-nois, USA
    3. Department of Genetics, Faculty of New Sciences and Technologies, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran


  • Introduction: Both eubacteria and archaea have an RNA-based defensive mechanism that can recognize and destroy external DNA and RNA. This provides acquired immunity from viral infections. An adaptive immune mechanism in prokaryotes known as CRISPR-associated (Cas) 9 enzyme (1–3) enables microorganisms to respond to and eliminate invading genetic material. (4). Based on numerous studies about the function of the CRISPR-Cas system, it has been widely used as a genome-editing tool. Due to its system's target specificity and ease of usage in targeting any gene of interest by simply altering the sequence of gRNA. (5) In addition to improving our understanding of complex biomolecular systems, this technology has also improved our abilities to take advantage of this process through techniques like genome editing, pathogen detection utilizing biosensors, and transcriptional regulation of genes (6). Furthermore, CRISPR/Cas-mediated identification of several microbial strains or detection of a disease with a single point mutation specificity has made this technique essential for diagnostic applications (7). Contrary to therapeutic applications, CRISPR diagnostic applications are often noninvasive and do not pose a threat to patient safety (8). This review article's goal is to provide an overview of molecular methods using the CRISPR-Cas system for identifying infectious ill-nesses.
  • Methods: Some ways have been utilized specifically to detect infectious diseases utilizing CRISPR-Cas in conjunction with other techniques, such as 1) The NASBACC, which is used to discriminate Zika virus strains. This technique combines NASBA with Cas9 cleavage activity (9). 2) CRISPR-MTB, a technique that could detect tuberculosis (10). 3) FLASH is a different CRISPR/Cas9-based technique designed to find pathogens that are resistant to medication, like Staphylococcus aureus and Plasmodium falciparum (11). (12). 4) COVID-19 has been identified by CRISPR-Dx (13). 5) A system called iSCAN uses CRISPR-Cas12 and RT-LAMP as effective COVID-19 detection modules (14). 6) DETECTR has been designed for the detection of SARS-CoV-2 and the human papillomavirus (HPV) (15, 16). 7) SARS-CoV-2 detection techniques have been cre-ated using CRISPR-FDS (17). 8) STOP (SHERLOCK Testing in One Pot), a straightforward test chemical for the one-hour detection of SARS-CoV-2, was developed (18). 9) A CREST platform has been developed using Cas13 detection in conjunction with a PCR, a linear amplifi-cation step (transcription), and enzymatic signal amplification via fluorescence detection to de-velop a cost-effective, sensitive, and user-friendly SARS-CoV-2 detection approach (19).
  • Results: Overall, in this study, we introduced several novel diagnostic tools that have primarily been uti-lized to identify infectious diseases and that primarily make use of Cas12 and Cas13 from the CRISPR complex. The majority of the aforementioned methods, such as CRISPR-Dx, iSCAN, DETECTR, CRISPR-FDS, STOP, and CREST, have been developed to identify SARS-CoV-2. These methods have two components, and CRISPR is an integral component of each one of them.
  • Conclusion: This research has presented a range of possible diagnostic methods for infectious disorders such as COVID-19, HPV, TB, and the Zika virus. In reality, efforts should be made to build these tools for the early and best identification of various infectious diseases with the aid of developed techniques like CRISPR.
  • Keywords: Diagnose, CRISPR-Cas, infectious, SARS-CoV-2