Hi everyone!
Is there a way to methylate the DNA of the living cells (in-vitro)?
For example, to demethylate the DNA we can use Decitabine. Is there a compound which initiates the opposite effect? Overexpression of Methyltransferase maybe?
Also, is it possible to do a targeted methylation? Say, methylate only a specific loci or CpG island of the genome?
Thank you!
Best,
Michail
DNMT1 enzyme can be used for targetted methylation of DNA. It catalyzes the transfer of methyl groups to specific CpG structures in DNA.
These may be of use:
Article DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A
https://www.nature.com/articles/ncomms16026
https://www.nature.com/articles/s41467-017-02708-5
What you ask has been a hot topic recently (especially with the introduction of Cas9-methyltransferase fusion enzymes). Petra has mentioned a few outstanding papers.
In addition, I suggest:
https://academic.oup.com/nar/article/45/4/1703/2605790
Article Targeted epigenetic editing of SPDEF reduces mucus productio...
Similarly, you can target demethylation using Cas9-TET fusions:
https://www.nature.com/articles/nbt.3658
For whole-genome hypermethylation you should look for papers on methyltransferase overexpression. Or TET protein knock-outs:
Article Tet1 and Tet2 Protect DNA Methylation Canyons against Hypermethylation
Using a recombinant methyltransferase, it is even possible to methylate a genome of a DNA methylation-free organism:
Article Cloning, characterization and expression of the gene coding ...
You can find much more papers to add to this list, just use appropriate keywords in search.
Dear all, thank you for your answers.
The usage of dCas9-MQ1 looks very promising to me!
Best,
Michail
And I am not aware of any chemical compound that would induce hypermethylation directly (like decitabine works for demethylation). Unless there is any TET inhibitor already developed, but upon a quick search I only found that "hypoxia causes DNA hypermethylation by reducing TET activity", no special drugs yet.
Article Tumor hypoxia causes DNA hypermethylation by reducing TET activity
To target CpG island no, but for studying the methylation levels of specific CpG Island try with this method.
https://academic.oup.com/biomethods/article/2/1/bpw008/2957405
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