Site-Specific Methylases

For a detailed, graphical explanation of the MaGellin work flow, please download the MaGellin Workflow Specifications Sheet, which includes all of the steps in the MaGellin workflow.

Now that the MaGellin assay was validated for non-specific methylases, we were ready to test site-specific methylases. Our MaGellin assay is ideal for high-throughput construction and testing of these enzymes. Site-specific methylases are fusion proteins, a DNA binding domain linked to a methylase by a serine glycine chain. They can direct DNA methylation to specific sequences, likely promoter regions for use as a transcriptional silencer. We used the prokaryotic methylase M.SssI for all of our studies (BBa_K1128000).

We wanted to first recapitulate published results with a zinc finger binding domain, and then characterize our two novel site-specific methylases: using the TALE and CRISPR/Cas binding domains.