Creating Custom DNA Binding Proteins
Author Information
Author(s): Weber Ernst, Gruetzner Ramona, Werner Stefan, Engler Carola, Marillonnet Sylvestre
Primary Institution: Icon Genetics GmbH, Halle/Saale, Germany
Hypothesis
Can we engineer TALE proteins with novel DNA binding specificities for genome engineering?
Conclusion
The study successfully demonstrates a method for assembling designer TALE proteins that can target specific DNA sequences in plants.
Supporting Evidence
- The study presents a strategy for engineering TALE proteins with novel DNA binding specificities.
- 68 repeat modules were created to allow for the assembly of DNA recognition domains for any 17 nucleotide target sequence.
- The Golden Gate cloning method was used for efficient assembly of multiple DNA fragments.
Takeaway
Scientists figured out how to make special proteins that can stick to specific parts of DNA, which can help in changing genes in plants.
Methodology
The study used a two-step Golden Gate cloning method to assemble TALE proteins with specific DNA binding domains.
Limitations
The cloning efficiency decreases when assembling 17 repeat modules in a single reaction.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website