Genome editing with GeneArt® Precision TALs





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Published on Aug 17, 2012

Learn more here: http://www.lifetechnologies.com/tal

Jon talks about the TAL effector proteins that were first identified as bacterial plant pathogens. Based on this, Life Technologies developed a new and powerful way to edit genomes and control gene function. The service GeneArt® Precision TALs provide custom DNA-binding proteins fused to a range of customer-designated effector domains that enable you to target the genome exactly where you want high precision and flexibility.


[Audio transcript:]

A new technology has been developed in the last few years that enables you to target the genome exactly where you want with high precision. These tools, called TAL effector proteins were originally identified as bacterial plant pathogens.
In nature TAL proteins are delivered to the plant host cell by the bacterium and are coded to bind to the genomic DNA at certain promoter elements, activating genes and facilitating infection.
Based on this technology we have developed a service that produces synthetically engineered versions of TAL binding proteins that enable you to target the genome exactly where you want.
Let's have a detailed look at this:
The GeneArt Precision TAL effectors are made up of two parts.
One part binds the DNA. You can design that to recognize just about any sequence you want. This binding part consists of 18 or 24 repeat modules. Each repeat is responsible for binding a specific base in the DNA and the repeats are not thought to interfere with each other. That allows us to engineer these molecules to bind any stretch of the DNA that you desire.
To the DNA binding part we fuse a second part: For example TALs with nuclease domains are used to generate double-stranded breaks by using two TAL effectors to each cleave a single strand. The space between the two individual TAL DNA binding sites is an important parameter for effective cleavage. Usually, a spacer of about 15 bp works well.
With this technique you can create gene knock-outs. The DNA ends formed by the clevage are repaired by the cell, often leaving deletions or small insertions. In these cases, two thirds of the repair events would alter the reading frame of a gene and create a knock-out.
Another option is to insert DNA sequences into the cut DNA. This process uses small ares of homology in the new 'donor' DNA to correct the DNA sequence or insert new DNA sequences. Ideally the insert sequence has overlapping homologous sequences on each side of about 300 to 500 bp.
We also offer an activator domain to control function of genes. That allows you to introduce these molecules into cells, whether it is a plant cell or an animal cell, and activate a gene specifically at the locus you want.
Additionally you can choose a repressor.
If you are looking at novel methods of genome modification and regulation and want to use your own effector you can choose the MCS vector and insert a protein-coding sequence of interest which allows you to create a TAL effector of your choice.
Examples for applications in both plant and animals can be; elucidating the basic function and regulation of a gene, studying metabolic pathways, embryonic stem cell research and research on disease models.
It is a powerful way to edit genomes and control gene function.
It's easy: Go to the TAL web page or into the GeneArt online order portal. Download the excel order form, fill in your binding sequences and effector choice and send the form to geneartsupport@lifetech.com
Within 2 weeks we will create a set of TAL effectors that bind to your specific target. The GeneArt Precision TALs are delivered it to you as Gateway® entry clones, ready for you to target the genome exactly where you want.

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