Team:Penn/FusionResults
From 2013.igem.org
Results
We ran our assay with the TALE Fusion, with or without the target site on the plasmid. We saw significantly different band patterns. The fusion did not cause off-target methylation when there was a target site included.
We quantified the gel with our MaGellin software
Advantages. Our new TALE fusion is also much more modular and easy to customize than the old zinc finger. TALE construction is not heavily patented like zinc-finger design, and TALE’s are considerably cheaper to produce (Sanjana 2012 and Zhang 2011). They have made the zinc-fingers nearly obsolete as a tool for genetic engineering, including the creation of transgenic animal models (Tesson 2011, Sander 2011, Huang 2011, and Zhang 2011). We expect our new fusion protein could be translated to enable the creation of differentially methylated animal models, which would be revolutionary for epigenetic disease research (Klose 2006). In effect, this could transform epigenetics from a largely observational discipline to one of active intervention and manipulation, similar to the transition from early classical genetics to genetic engineering and synthetic biology.
Our TALE fusion has numerous advantages over the existing Zinc Finger fusions.
- We have developed a novel TALE-M.SssI fusion protein that works better than existing equivalents
- Our protein is more specific
- TALE binding domains are easy to customize and tailor to target specific sequences at lower costs
- Synthetic biologists could use our protein to begin experimenting with an orthogonal level of control in E. coli – they can begin to engineer the epigenome