Dataset Information

Miniaturization of CRISPR/Cas12-Based DNA Sensor Array by Non-Contact Printing.

ABSTRACT: DNA microarrays have been applied for comprehensive genotyping, but remain a drawback in complicated operations. As a solution, we previously reported the solid-phase collateral cleavage (SPCC) system based on the clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 12 (CRISPR/Cas12). Surface-immobilized Cas12-CRISPR RNA (crRNA) can directly hybridize target double-stranded DNA (dsDNA) and subsequently produce a signal via the cleavage of single-stranded DNA (ssDNA) reporter immobilized on the same spot. Therefore, SPCC-based multiplex dsDNA detection can be performed easily. This study reports the miniaturization of SPCC-based spots patterned by a non-contact printer and its performance in comprehensive genotyping on a massively accumulated array. Initially, printing, immobilization, and washing processes of Cas12-crRNA were established to fabricate the non-contact-patterned SPCC-based sensor array. A target dsDNA concentration response was obtained based on the developed sensor array, even with a spot diameter of 0.64 ± 0.05 mm. Also, the limit of detection was 572 pM, 531 pM, and 3.04 nM with 40, 20, and 10 nL-printing of Cas12-crRNA, respectively. Furthermore, the sensor array specifically detected three dsDNA sequences in one-pot multiplexing; therefore, the feasibility of comprehensive genotyping was confirmed. These results demonstrate that our technology can be miniaturized as a CRISPR/Cas12-based microarray by using non-contact printing. In the future, the non-contact-patterned SPCC-based sensor array can be applied as an alternative tool to DNA microarrays.

SUBMITTER: Shigemori H

PROVIDER: S-EPMC10818962 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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Miniaturization of CRISPR/Cas12-Based DNA Sensor Array by Non-Contact Printing.

Shigemori Hiroki H Fujita Satoshi S Tamiya Eiichi E Nagai Hidenori H

Micromachines 20240117 1

DNA microarrays have been applied for comprehensive genotyping, but remain a drawback in complicated operations. As a solution, we previously reported the solid-phase collateral cleavage (SPCC) system based on the clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 12 (CRISPR/Cas12). Surface-immobilized Cas12-CRISPR RNA (crRNA) can directly hybridize target double-stranded DNA (dsDNA) and subsequently produce a signal via the cleavage of single-stranded DNA (ssDNA) ...[more]

PMID: 38258263

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Project description:It has been the dream of many scientists and engineers to realize a non-contact remote sensing system that can perform continuous, accurate and long-term monitoring of human vital signs as we have seen in many Sci-Fi movies. Having an intelligible sensor system that can measure and record key vital signs (such as heart rates and respiration rates) remotely and continuously without touching the patients, for example, can be an invaluable tool for physicians who need to make rapid life-and-death decisions. Such a sensor system can also effectively help physicians and patients making better informed decisions when patients' long-term vital signs data is available. Therefore, there has been a lot of research activities on developing a non-contact sensor system that can monitor a patient's vital signs and quickly transmit the information to healthcare professionals. Doppler-based radio-frequency (RF) non-contact vital signs (NCVS) monitoring system are particularly attractive for long term vital signs monitoring because there are no wires, electrodes, wearable devices, nor any contact-based sensors involved so the subjects may not be even aware of the ubiquitous monitoring. In this paper, we will provide a brief review on some latest development on NCVS sensors and compare them against a few novel and intelligent phased-array Doppler-based RF NCVS biosensors we have built in our labs. Some of our NCVS sensor tests were performed within a clutter-free anechoic chamber to mitigate the environmental clutters, while most tests were conducted within the typical Herman-Miller type office cubicle setting to mimic a more practical monitoring environment. Additionally, we will show the measurement data to demonstrate the feasibility of long-term NCVS monitoring. The measured data strongly suggests that our latest phased array NCVS system should be able to perform long-term vital signs monitoring intelligently and robustly, especially for situations where the subject is sleeping without hectic movements nearby.

Dataset Information

Miniaturization of CRISPR/Cas12-Based DNA Sensor Array by Non-Contact Printing.

Publications

Miniaturization of CRISPR/Cas12-Based DNA Sensor Array by Non-Contact Printing.

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