PMID- 38280781
OWN - NLM
STAT- MEDLINE
DCOM- 20240129
LR  - 20240129
IS  - 1873-4324 (Electronic)
IS  - 0003-2670 (Linking)
VI  - 1291
DP  - 2024 Feb 22
TI  - Leveraging CRISPR/Cas12 signal amplifier to sensitive detection of 
      apurinic/apyrimidinic endonuclease 1 and high-throughput inhibitor screening.
PG  - 342212
LID - S0003-2670(24)00013-8 [pii]
LID - 10.1016/j.aca.2024.342212 [doi]
AB  - As an essential protein in DNA repair, apurinic/apyrimidinic endonuclease 1 
      (APE1) plays multiple critical functions in maintaining homeostasis, making it a 
      significant biomarker and therapeutic target for many disorders. Here, we 
      describe a simple method to detect APE1 based on the Releasing-Extension-Signal 
      amplification Test (REST) strategy that leverages the dsDNA as the activator to 
      fully unlock the trans-cleavage activity of CRISPR/Cas12a. This assay provides a 
      rapid and specific APE1 detection with a detection limit down to 1.05 x 10(-5) 
      U/mL. We also combined this method with an automated pipetting platform and a 
      microplate reader for high-throughput screening of potential inhibitors of APE1. 
      Besides, by changing the modification on the probe, the REST strategy was easily 
      repurposed to detect various DNA glycosylases. Taken together, the simplicity and 
      robustness of the method offer a new choice for APE1 detection and inhibitor 
      screening, showing great potential in practical use. Furthermore, the REST 
      strategy devised in this study provides a new example of applying CRISPR/Cas12a 
      signal amplifier to non-nucleic acid biosensing and inhibitor screening, which 
      broadens the CRISPR-Dx toolbox.
CI  - Copyright (c) 2024 Elsevier B.V. All rights reserved.
FAU - Ding, Sheng
AU  - Ding S
AD  - Clinical Medical College & Affiliated Hospital, Chengdu University, Chengdu, 
      610041, PR China.
FAU - Yuan, Yi
AU  - Yuan Y
AD  - Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy 
      of Science, Chengdu, 610041, PR China.
FAU - Dong, Juan
AU  - Dong J
AD  - Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy 
      of Science, Chengdu, 610041, PR China.
FAU - Du, Feng
AU  - Du F
AD  - Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy 
      of Science, Chengdu, 610041, PR China. Electronic address: dufeng@cib.ac.cn.
FAU - Cui, Xin
AU  - Cui X
AD  - Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy 
      of Science, Chengdu, 610041, PR China.
FAU - Shi, Zheng
AU  - Shi Z
AD  - Clinical Medical College & Affiliated Hospital, Chengdu University, Chengdu, 
      610041, PR China.
FAU - Tang, Zhuo
AU  - Tang Z
AD  - Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy 
      of Science, Chengdu, 610041, PR China. Electronic address: tangzhuo@cib.ac.cn.
LA  - eng
PT  - Journal Article
DEP - 20240103
PL  - Netherlands
TA  - Anal Chim Acta
JT  - Analytica chimica acta
JID - 0370534
RN  - EC 4.2.99.18 (DNA-(Apurinic or Apyrimidinic Site) Lyase)
RN  - EC 3.1.- (Endonucleases)
SB  - IM
MH  - *High-Throughput Screening Assays
MH  - *CRISPR-Cas Systems
MH  - DNA Repair
MH  - DNA-(Apurinic or Apyrimidinic Site) Lyase/metabolism
MH  - Endonucleases/metabolism
OTO - NOTNLM
OT  - APE1
OT  - CRISPR/Cas12a
OT  - High-throughput inhibitor screening
OT  - Non-nucleic acid biosensing
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2024/01/28 07:44
MHDA- 2024/01/29 06:43
CRDT- 2024/01/27 20:58
PHST- 2023/11/12 00:00 [received]
PHST- 2024/01/01 00:00 [revised]
PHST- 2024/01/02 00:00 [accepted]
PHST- 2024/01/29 06:43 [medline]
PHST- 2024/01/28 07:44 [pubmed]
PHST- 2024/01/27 20:58 [entrez]
AID - S0003-2670(24)00013-8 [pii]
AID - 10.1016/j.aca.2024.342212 [doi]
PST - ppublish
SO  - Anal Chim Acta. 2024 Feb 22;1291:342212. doi: 10.1016/j.aca.2024.342212. Epub 
      2024 Jan 3.