Penn iGEM

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.

After validating the MaGellin assay with 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 a novel site-specific methylase using a TALE DNA binding domain.

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