The application of DNA polymerases and Cas9 as representative of DNA-modifying enzymes group in DNA sensor design (review)

Biosens Bioelectron. 2021 Mar 1:175:112867. doi: 10.1016/j.bios.2020.112867. Epub 2020 Dec 3.

Abstract

Rapid detection of nucleic acids (DNA or RNA) by inexpensive, selective, accurate, and highly sensitive methods is very important for biosensors. DNA-sensors based on DNA-modifying enzymes for fast determination and monitoring of pathogenic (Zika, Dengue, SARS-Cov-2 (inducer of COVID-19), human papillomavirus, HIV, etc.) viruses and diagnosis of virus-induced diseases is a key factor of this overview. Recently, DNA-modifying enzymes (Taq DNA polymerase, Phi29 DNA polymerase) have been widely used for the diagnosis of virus or pathogenic disease by gold standard (PCR, qPCR, RT-qPCR) methods, therefore, alternative methods have been reviewed. The main mechanisms of DNA metabolism (replication cycle, amplification) and the genomeediting tool CRISPR-Cas9 are purposefully discussed in order to address strategic possibility to design DNA-sensors based on immobilized DNA-enzymes. However, the immobilization of biologically active proteins on a gold carrier technique with the ability to detect viral or bacterial nucleic acids is individual for each DNA-modifying enzyme group, due to a different number of active sites, C and N terminal locations and arrangement, therefore, individual protocols based on the 'masking' of active sites should be elaborated for each enzyme.

Keywords: Biosensors; COVID-19; CRISPR-Cas9; DNA polymerases; DNA-Sensors; Gold nanoparticles.

Publication types

  • Review

MeSH terms

  • Biosensing Techniques*
  • COVID-19 / diagnosis*
  • COVID-19 / genetics
  • COVID-19 / virology
  • CRISPR-Cas Systems / genetics
  • Diagnostic Tests, Routine
  • Humans
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification*
  • SARS-CoV-2 / isolation & purification*
  • SARS-CoV-2 / pathogenicity

Substances

  • RNA, Viral