Journal
BIOSENSORS & BIOELECTRONICS
Volume 173, Issue -, Pages -Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2020.112619
Keywords
Transcription factors; CRISPR; Cas12a; Biosensors; DNA binding proteins
Categories
Funding
- Natural Science Foundation of Jiangsu Province [BK20200718, BK20200728]
- National Natural Science Foundation of China [22007049]
- Natural Science Foundation of the Jiangsu Higher Education Institutions of China
- Scientific Research Start-Up Funding for Nanjing Normal University [184080H202B246]
- Postgraduate Research & Practice Innovation Program of Jiangsu Province [KYCX20_1261]
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Transcription factors (TFs) are crucial proteins for cell fate determination and have potential applications in disease diagnostics and treatment. A highly sensitive biosensor based on the CRISPR/Cas12a system was developed for TF detection, utilizing competitive binding between Cas12a-crRNA and TFs to a specific dsDNA activator. This study demonstrates a novel approach in applying CRISPR technology for TF sensing and highlights the potential of using blocking mechanisms as a new sensing strategy for CRISPR-based biosensors.
Transcription factors (TFs) are the key proteins for the decision of cell fates, and they have been recognized as potent markers for diagnostic and treatment of diseases. Herein, we report on a highly sensitive biosensor for the detection of TFs based on the CRISPR/Cas12a system. This biosensor was accomplished based on the competitive binding of the Cas12a-crRNA and TFs towards a dsDNA referred to as activator. Without TFs, the activator can be recognized by Cas12a-crRNA and cause the activation of the DNase activity of Cas12a. When TFs were added, the TFs can bind with the activator because the activator was designed to contain the specific binding sites of target TFs. We find that this binding can inhibit the association between Cas12a-crRNA and the activator, which hinders the activation of Cas12a. As a proof-of-concept, the rapid detection of five kinds of TFs was presented, and the detection was extended to the analysis of TFs expression in xenograft solid tumors from mice. This investigation is the first attempt to apply CRISPR technology in the sensing of TFs, and it discloses that the blocking of activator can be applied as a new sensing mechanism for the development of CRISPR-based biosensor.
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