Hello,
I am a relatively new in DNA methylation field. I am interested in DNA methylation status of some CpGs sites: two from exons and one from introne. I would like to monitor their methylation status without killing the cells, so conventional sequencing approach do not work. I found the so-called Reporter of Genomic Methylation (RGM) that relies on a minimal imprinted gene promoter driving a fluorescent protein (Tracing Dynamic Changes of DNA Methylation at Single-Cell Resolution https://www.cell.com/cell/references/S0092-8674(15)01098-3 ).
Can someone please explain will the following work or not and why? I plan to make AAV vector with 100-200 bp with the CpG of interest + RGM. These bp also have additional methylated-capable CpGs which correlate with methylation status of the CpG of interest, so if it will report the others it is also fine.
I will appreciate every suggestions or references
Thank you,
Nikita
Hi colleague
Find the following URL may help you:
Article Tracing Dynamic Changes of DNA Methylation at Single-Cell Resolution
Regards...
Prof. Ayad Ghany Ismaeel , Thank you for the answer. This the paper, which I mentioned in the question. In the article, they provide examples with superenhancers, which are distal regions from coding regions in DNA, as well as with a promoter + the reporter. Both are not coding regions and will not interfere with protein expression. In my case, I am interested in regions which are inside of coding regions. Maybe, if I will add the reporter from the article in protein-coding region, it will interfere with protein expression and will make cells not viable. Thus, I think about a similar approach: to take a region of interest and the reporter downstream and insert it into genome.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line