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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.