CRISPR-HOLMES-based NAD⁺ detection

Zhuang, Songkuan1,2; Hu, Tianshuai2; Zhou, Hongzhong2; He, Shiping2; Li, Jie2; Zhang, Yuehui3; Gu, Dayong2; Xu, Yong4; Chen, Yijian5,6; Wang, Jin1,2,7

2024-03-25

10.3389/fbioe.2024.1355640

FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY   影响因子和分区

2296-4185

Studies have indicated that the intracellular nicotinamide adenine dinucleotide (NAD+) level is associated with the occurrence and development of many diseases. However, traditional nicotinamide adenine dinucleotide (NAD+) detection techniques are time-consuming and may require large and expensive instruments. We recently found that the clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a protein can be inactivated by AcrVA5-mediated acetylation and reactivated by CobB, using NAD+ as the co-factor. Therefore, in this study, we created a CRISPR-Cas12a-based one-step HOLMES(NAD+) system for rapid and convenient NAD+ detection with the employment of both acetylated Cas12a and CobB. In HOLMES(NAD+), acetylated Cas12a loses its trans-cleavage activities and can be reactivated by CobB in the presence of NAD+, cutting ssDNA reporters to generate fluorescence signals. HOLMES(NAD+) shows both sensitivity and specificity in NAD+ detection and can be used for quantitative determination of intracellular NAD+ concentrations. Therefore, HOLMES(NAD+) not only provides a convenient and rapid approach for target NAD+ quantitation but also expands the application scenarios of HOLMES to non-nucleic acid detection.

CRISPR Cas12a HOLMES NAD(+) acetylation

SCI ; EI

英语

通讯

Biotechnology & Applied Microbiology ; Engineering

Biotechnology & Applied Microbiology ; Engineering, Biomedical

WOS:001198054700001

FRONTIERS MEDIA SA

Web of Science

1.Shenzhen Univ, Sch Biomed Engn, Guangdong Key Lab Biomed Measurements & Ultrasound, Natl Reg Key Technol Engn Lab Med Ultrasound,Med S, Shenzhen, Peoples R China
2.Shenzhen Univ, Shenzhen Peoples Hosp 2, Shenzhen Inst Translat Med, Dept Clin Lab,Affiliated Hosp 1, Shenzhen, Peoples R China
3.Shenzhen Bao An Peoples Hosp, Shenzhen, Peoples R China
4.Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Affiliated Hosp 2, Natl Clin Res Ctr Infect Dis,Dept Clin Lab, Shenzhen, Peoples R China
5.Fudan Univ, Huashan Hosp, Inst Antibiot, Shanghai, Peoples R China
6.Natl Hlth Commiss, Key Lab Clin Pharmacol Antibiot, Shanghai, Peoples R China
7.Shanghai Tolo Biotechnol Co Ltd, Shanghai, Peoples R China

Zhuang, Songkuan,Hu, Tianshuai,Zhou, Hongzhong,et al. CRISPR-HOLMES-based NAD⁺ detection[J]. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY,2024,12.

Zhuang, Songkuan.,Hu, Tianshuai.,Zhou, Hongzhong.,He, Shiping.,Li, Jie.,...&Wang, Jin.(2024).CRISPR-HOLMES-based NAD⁺ detection.FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY,12.

Zhuang, Songkuan,et al."CRISPR-HOLMES-based NAD⁺ detection".FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY 12(2024).