Protocol Library

Find and share protocols to run on your OT-One


Developed by: opentrons

The FusX Automated TALEN Kit is capable of building over 100 TALEN coding plasmids with no need for additional synthesized parts. Now labs can edit any genomic DNA sequence they wish from their own bench -- no more relying on external oligo synthesis to do your genome editing experiments.

The custom TALEN plasmid is made by ligating six DNA parts into a final receiver plasmid via Golden Gate assembly. In this single tube reaction a combination of FusX plasmids containing two or three RVD cassettes are bundled together. The FusX kit includes every possible RVD combination that can be made. These components - FusX plasmids plus reciever plasmids - make up the 343 unique FusX parts that can be combined to code for any desired TALEN.

Our FusX Protocol Generator and the OT-One simplify the complex combinatorial step of putting together the correct FusX parts for each Golden Gate assembly. Users input the DNA target sequence to the generator, and the back-end software determines which FusX parts to assemble and automatically generates an OT-One protocol. This protocol, when run on an OT-One with the FusX Parts Plates on deck, results in a custom FusX plasmid mix ready to go into the thermocycler for Golden Gate assembly.

Estimated time for this protocol is 12 minutes (1 TALEN)

Setup topTop Icon

    • A1 trash
    • B1 p10-rack1
    • B2 p10-rack2
    • B3 p10-rack3
    • C1 TALE1
    • C2 TALE3
    • C3 TALE5
    • D1 TALE2
    • D2 TALE4
    • E1 FusX Output
    • E3 Ingredients
    • 12 minutes (1 TALEN)
    • 22 minutes (2 TALENS)
    • 32 minutes (3 TALENS)
    • 9 x p10 tips (1 TALEN)
    • 17 x p10 tips (2 TALENs)
    • 25 x p10 tips (3 TALENs)

Click on each individual container to zoom in.

Fusx Protocol Generatortop

The protocol generator can create up to 15 unique sequences in one run, and can take either an RVD or DNA input (spaces are ignored). Please enter each sequence on its own line. Once your protocol has been created, final well positions for each target sequence will be shown below.

Example DNA: A C T T G C T A G G C A A T C


FusX Output Plate:

Information topTop Icon

The Lifengine FusX Kit is the world’s first all-inclusive, inhouse, automated gene editor production system. Input your DNA target sequence to Mix.Bio, place the FusX Part Plates on your OT-One, and hit ‘run’ to construct your custom TALEN.

The Lifengine FusX kit based on the pFusX system developed at the Mayo Clinic and published in the Journal of Human Gene Therapy.

Genome engineering is biological reprogramming. It is the process of making defined changes in DNA, the software of the cell. Modern genome editing tools focus on the ability to target unique sequences of DNA with a ‘cursor’ that hones in on the right part of the genome to label, modify, or break DNA in very specific places. One DNA ‘cursor’ tool is the Transcription Activator-Like Effector (TALE), a customizable DNA binding protein originally discovered in pathogenic plant bacteria. TALEs rely on a simple mechanism of amino acid sequences containing Repeat Variable Diresidues (RVDs) to identify and bind to any target DNA sequence. Each RVD recognizes one DNA basepair, and combining RVDs into a string creates the code for a TALE’s target sequence.

The TALE ‘cursor’ proteins can be paired with a nuclease (N) protein, which cleaves both strands of DNA. The TALE’s programmed RVDs affix themselves to the corresonding base-pair section of target genomic DNA to guide the Nuclease to the right cleavage point. These target-and-cut protein combos are called TALENs. TALENs work in pairs, flanking the sequence of DNA they want to cut like two scissor blades coming together. Because of their exceptional cutting ability and high precision TALENs are one of the most trusted and flexible genome engineering tools in use today.

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