Sequence-Specific Recognition of HIV-1 DNA with Solid-State CRISPR-Cas12a-Assisted Nanopores (SCAN).
Reza NouriYuqian JiangXiaojun Lance LianWeihua GuanPublished in: ACS sensors (2020)
Nucleic acid detection methods are crucial for many fields such as pathogen detection and genotyping. Solid-state nanopore sensors represent a promising platform for nucleic acid detection due to its unique single molecule sensitivity and label-free electronic sensing. Here, we demonstrated the use of the glass nanopore for highly sensitive quantification of single-stranded circular DNAs (reporters), which could be degraded under the trans-cleavage activity of the target-specific CRISPR-Cas12a. We developed and optimized the Cas12a assay for HIV-1 analysis. We validated the concept of the solid-state CRISPR-Cas12a-assisted nanopores (SCAN) to specifically detect the HIV-1 DNAs. We showed that the glass nanopore sensor is effective in monitoring the cleavage activity of the target DNA-activated Cas12a. We developed a model to predict the total experimental time needed for making a statistically confident positive/negative call in a qualitative test. The SCAN concept combines the much-needed specificity and sensitivity into a single platform, and we anticipate that the SCAN would provide a compact, rapid, and low-cost method for nucleic acid detection at the point of care.
Keyphrases
- solid state
- nucleic acid
- crispr cas
- label free
- genome editing
- single molecule
- loop mediated isothermal amplification
- low cost
- antiretroviral therapy
- computed tomography
- hiv positive
- hiv infected
- human immunodeficiency virus
- hiv testing
- high throughput
- hepatitis c virus
- hiv aids
- men who have sex with men
- living cells
- genome wide
- gene expression
- magnetic resonance imaging
- dna methylation
- single cell
- cell free
- mass spectrometry
- south africa
- high resolution
- amino acid
- fluorescent probe
- simultaneous determination