2. Academic Validation
  3. A CRISPR-derived biosensor for the sensitive detection of transcription factors based on the target-induced inhibition of Cas12a activation

A CRISPR-derived biosensor for the sensitive detection of transcription factors based on the target-induced inhibition of Cas12a activation

  • Biosens Bioelectron. 2020 Sep 19;173:112619. doi: 10.1016/j.bios.2020.112619.
Bingzhi Li 1,  Anqi Xia 1,  Shilin Zhang 1,  Tiying Suo 1,  Yujie Ma 2,  He Huang 3,  Xing Zhang 4,  Yue Chen 5,  Xuemin Zhou 6
Affiliations

Affiliations

  • 1 School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, China.
  • 2 School of Nursing, Nanjing Medical University, Nanjing, 211166, China.
  • 3 School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, China. Electronic address: huangh@njnu.edu.cn.
  • 4 School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, China. Electronic address: zhangxing@njnu.edu.cn.
  • 5 School of Nursing, Nanjing Medical University, Nanjing, 211166, China. Electronic address: chenyue1005@njmu.edu.cn.
  • 6 School of Pharmacy, Nanjing Medical University, Nanjing, 211166, China. Electronic address: xueminzhou001_001@hotmail.com.
PMID: 33221511 DOI: 10.1016/j.bios.2020.112619
Abstract

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.

Keywords

Biosensors; CRISPR; Cas12a; DNA binding proteins; Transcription factors.

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