meHOLMES: A CRISPR-cas12a-based method for rapid detection of DNA methylation in a sequence-independent manner

Zhuang，Songkuan1,2; Hu，Tianshuai1; Zhou，Xike3; Zhou，Hongzhong1; He，Shiping1; Li，Jie1; Qiu，Long6; Zhang，Yuehui4; Xu，Yong5; Pei，Hao3; Gu，Dayong1; Wang，Jin1,6

2024-01-30

10.1016/j.heliyon.2024.e24574

Heliyon   影响因子和分区

2405-8440

Aberrant DNA methylation is closely associated with various diseases, particularly cancer, and its precise detection plays an essential role in disease diagnosis and monitoring. In this study, we present a novel DNA methylation detection method (namely meHOLMES), which integrates both the TET2/APOBEC-mediated cytosine deamination step and the CRISPR-Cas12a-based signal readout step. TET2/APOBEC efficiently converts unmethylated cytosine to uracil, which is subsequently changed to thymine after PCR amplification. Utilizing a rationally designed crRNA, Cas12a specifically identifies unconverted methylated cytosines and generates detectable signals using either fluorescent reporters or lateral flow test strips. meHOLMES quantitatively detects methylated CpG sites with or without Protospacer Adjacent Motif (PAM) sequences in both artificial and real biological samples. In addition, meHOLMES can complete the whole detection process within 6 h, which is much faster than traditional bisulfite-based sample pre-treatment method. Above all, meHOLMES provides a simpler, faster, more accurate, and cost-effective approach for quantitation of DNA methylation levels in a sequence-independent manner.

APOBEC CRISPR-Cas12a DNA methylation HOLMES TET2

SCI

英语

其他

2-s2.0-85183203528

Scopus

1.Department of Clinical Laboratory,Shenzhen Institute of Translational Medicine,The First Affiliated Hospital of Shenzhen University,Shenzhen Second People's Hospital,Shenzhen,518060,China
2.Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging,National-Regional Key Technology Engineering Laboratory for Medical Ultrasound,School of Biomedical Engineering,Shenzhen University Medical School,Shenzhen,518060,China
3.The Fifth People's Hospital of Wuxi,Affiliated to Jiangnan University,Wuxi,Jiangsu,214007,China
4.Shenzhen Bao An Peoples Hospital,Shenzhen,518060,China
5.Department of Clinical Laboratory,Shenzhen Third People's Hospital,The Second Affiliated Hospital,School of Medicine,Southern University of Science and Technology,National Clinical Research Center for Infectious Disease,Shenzhen,518112,China
6.Tolo Biotechnology Co.,Ltd,Wuxi,Jiangsu,214174,China

Zhuang，Songkuan,Hu，Tianshuai,Zhou，Xike,et al. meHOLMES: A CRISPR-cas12a-based method for rapid detection of DNA methylation in a sequence-independent manner[J]. Heliyon,2024,10(2).

Zhuang，Songkuan.,Hu，Tianshuai.,Zhou，Xike.,Zhou，Hongzhong.,He，Shiping.,...&Wang，Jin.(2024).meHOLMES: A CRISPR-cas12a-based method for rapid detection of DNA methylation in a sequence-independent manner.Heliyon,10(2).

Zhuang，Songkuan,et al."meHOLMES: A CRISPR-cas12a-based method for rapid detection of DNA methylation in a sequence-independent manner".Heliyon 10.2(2024).