Precise Recognition and Quantification of Locus‐Specific DNA Methylation Using Engineered ROS1† DOI
Li Zeng,

Fang‐Yin Gang,

Tong‐Tong Ji

et al.

Chinese Journal of Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 24, 2025

Comprehensive Summary 5‐Methylcytosine (5mC) plays pivotal roles in numerous biological processes. To gain a deeper understanding of the functions 5mC, it is essential to develop methods for its quantitative analysis. Herein, we engineered repressor silencing 1 (ROS1) protein enhance glycosylase/lyase activity towards resulting an ROS1 (eROS1) that can effectively excise 5mC from DNA. Using eROS1, developed method termed ROS1‐mediated (eRMQ) analysis, locus‐specific quantification genomic This capitalizes on ability eROS1 selectively cleave which creates one‐nucleotide gap. The presence this gap hinders extension DNA polymerase, leading reduction products be evaluated using real‐time PCR (qPCR). limit detection eRMQ was as low fM. method, achieved analysis at individual sites within and demonstrated significant levels lung cancer tissues compared adjacent normal tissues. Taken together, study introduces DNA, offering valuable tool exploring epigenetic regulation human diseases.

Language: Английский

Recent advance in single-molecule detection and imaging DOI
Han Yun,

Weijie Tong,

Fei Ma

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 118252 - 118252

Published: April 1, 2025

Language: Английский

Citations

0
Direct Single-Nucleotide Resolution Sequencing of DNA 5-Methylcytosine Using Engineered DNA Methyltransferase-Mediated CMD-seq DOI Creative Commons
Wei Liu, Zhaocheng Ma, Shan Zhang

et al.

Chemical Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Utilizing the newly identified M.MedI-N377K methyltransferase, we developed a novel method of CMD-seq for stoichiometric detection 5mC in DNA at single-nucleotide resolution.

Language: Английский

Citations

0
Precise Recognition and Quantification of Locus‐Specific DNA Methylation Using Engineered ROS1† DOI
Li Zeng,

Fang‐Yin Gang,

Tong‐Tong Ji

et al.

Chinese Journal of Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 24, 2025

Comprehensive Summary 5‐Methylcytosine (5mC) plays pivotal roles in numerous biological processes. To gain a deeper understanding of the functions 5mC, it is essential to develop methods for its quantitative analysis. Herein, we engineered repressor silencing 1 (ROS1) protein enhance glycosylase/lyase activity towards resulting an ROS1 (eROS1) that can effectively excise 5mC from DNA. Using eROS1, developed method termed ROS1‐mediated (eRMQ) analysis, locus‐specific quantification genomic This capitalizes on ability eROS1 selectively cleave which creates one‐nucleotide gap. The presence this gap hinders extension DNA polymerase, leading reduction products be evaluated using real‐time PCR (qPCR). limit detection eRMQ was as low fM. method, achieved analysis at individual sites within and demonstrated significant levels lung cancer tissues compared adjacent normal tissues. Taken together, study introduces DNA, offering valuable tool exploring epigenetic regulation human diseases.

Language: Английский

Citations

0