Accurate and efficient detection of DNA is crucial for disease diagnosis and health monitoring. The traditional methods for DNA analysis involve multiple steps, including sample preparation, lysis, extraction, amplification, and detection. In this study, we present a one-step elution-free DNA analysis method based on the combination of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated light-up aptamer transcription (CLAT) assay and a DNA-capturing poly(2-dimethylaminomethyl styrene) (pDMAMS)-coated tube. The sample solution and lysis buffer are added to the pDMAMS-coated tube, and the DNA is efficiently captured on the surface via electrostatic interaction and directly detected by CLAT assay. The ability of the CRISPR/Cas9 system to specifically recognize DNA enables direct detection of DNA captured on the pDMAMS-coated tube. The combination of CLAT assay and pDMAMS-coated tube simplifies DNA detection in a single tube without the need for complicated extraction steps, improving sensitivity. Our platform demonstrated attomolar sensitivity in the detection of target DNA in cell lysate (0.92 aM), urine (7.7 aM), and plasma (94.6 aM) samples within 1 h. The practical applicability of this method was further demonstrated in experiments with tumor-bearing mice. We believe that this approach brings us closer to an all-in-one DNA purification and detection tube system and has potential applications in tissue and liquid biopsies, as well as various other DNA sensing applications.
|Journal||Biosensors and Bioelectronics|
|Publication status||Published - 2023 Apr 1|
Bibliographical noteFunding Information:
This research was supported by National Research Foundation ( NRF ) grants funded by Korea government ( MSIT ) ( NRF-2021M3E5E3080379 , NRF-2021M3H4A1A02051048 , NRF-2021M3E5E3080844 , NRF-2022R1C1C1008815 , NRF-2020R1A2C1010453 , NRF-2021M3H4A4079293 , and NRF-2021R1A2B5B03001416 ), Technology Development Program for Biological Hazards Management in Indoor Air through Korea Environment Industry & Technology Institute ( KEITI ) funded by Korea government ( ME ) ( 2021003370003 ), Korea Evaluation Institute of Industrial Technology ( KEIT ) grant funded by Korea government ( MOTIE ) ( RS-2022-00154853 ), Nanomedical Devices Development Program of National Nano Fab Center (CSM2105M101 and CSM2102M101), and KRIBB Research Initiative Program ( 1711134081 ).
© 2023 The Authors
All Science Journal Classification (ASJC) codes
- Biomedical Engineering