A Methyl-Engineered DNAzyme for Endogenous Alkyltransferase Monitoring and Self-Sufficient Gene Regulation.
Yuqiu HeQing WangQingqing ZhangYifei WangYuqian JiangQiu ZhaoXiaoqing LiuFuan WangPublished in: Small methods (2024)
The on-demand gene regulation is crucial for extensively exploring specific gene functions and developing personalized gene therapeutics, which shows great promise in precision medicines. Although some nucleic acid-based gene regulatory tools (antisense oligonucleotides and small interfering RNAs) are devised for achieving on-demand activation, the introduction of chemical modifications may cause undesired side effects, thereby impairing the gene regulatory efficacy. Herein, a methyl-engineered DNAzyme (MeDz) is developed for the visualization of endogenous alkyltransferase (AGT) and the simultaneous self-sufficiently on-demand gene regulation. The catalytic activity of DNAzyme can be efficiently blocked by O 6 -methylguanine (O 6 MeG) modification and specifically restored via the AGT-mediated DNA-repairing pathway. This simply designed MeDz is demonstrated to reveal AGT of varying expression levels in different cells, opening the possibility to explore the AGT-related biological processes. Moreover, the AGT-guided MeDz exhibits cell-selective regulation on the human early growth response-1 (EGR-1) gene, with efficient gene repression in breast cancer cells and low effectiveness in normal cells. The proposed MeDz offers an attractive strategy for on-demand gene regulation, displaying great potential in biomedical applications.
Keyphrases
- nucleic acid
- genome wide
- induced apoptosis
- copy number
- cell cycle arrest
- breast cancer cells
- genome wide identification
- living cells
- endothelial cells
- single cell
- systematic review
- cell death
- endoplasmic reticulum stress
- dna methylation
- signaling pathway
- multidrug resistant
- cell proliferation
- transcription factor
- resting state
- functional connectivity
- big data
- fluorescent probe
- circulating tumor cells