Talens
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07:42
Genome Editing in Astyanax mexicanus Using Transcription Activator-like Effector Nucleases (TALENs)
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Cited by 21 •
2016
Gene-targeting mutagenesis is now possible in a wide range of organisms using genome editing techniques. Here, we demonstrate a protocol for targeted gene mutagenesis using transcription activator like effector nucleases (TALENs) in Astyanax mexicanus, a species of fish that includes surface fish and cavefish.
04:24
TALEN-Mediated Targeted Gene Integration: Using Engineered Sequence-Specific Nucleases for the Precise Insertion of Fluorescent Protein Gene into hiPSCs
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2025
This video describes a precise genome editing technique in human induced pluripotent stem cells, or hiPSCs, using TALENs to create double-stranded breaks at a targeted locus, inducing homology-directed repair for fluorescence protein gene integration.
07:28
Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
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Cited by 12 •
2015
TALEN-mediated gene editing at the safe harbor AAVS1 locus enables high-efficiency transgene addition in human iPSCs. This protocol describes the procedures for preparing iPSCs for TALEN and donor vector delivery, transfecting iPSCs, and selecting and isolating iPSC clones to achieve targeted integration of a GFP gene to generate reporter lines.
08:31
Genome Editing
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2023
A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target genomic sites by either proteins or RNAs that recognize specific sequences. When a cell attempts to repair this damage, mutations can be...
12:04
Mouse Genome Engineering Using Designer Nucleases
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Cited by 14 •
2014
Designer nucleases such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be used to modify the genome of mouse preimplantation embryos by triggering both the nonhomologous end joining (NHEJ) and homologous recombination (HR) pathways. These advances enable the rapid generation of mice with precise genetic modifications.
FAQs
Specificity comes from the DNA-binding portion of each engineered protein. Customizable TALE repeats are arranged to recognize selected nucleotide sequences, so a matched pair binds opposite DNA strands at the intended site. Cleavage occurs only after the two bound proteins bring their FokI domains together. This paired requirement connects sequence recognition with controlled cutting.
FokI provides the cutting activity, but it does not act as an isolated TALEN component at the target. The nuclease domains must dimerize after both TALENs bind their respective DNA sites. Consequently, successful editing depends on coordinated recognition of two nearby, opposite-strand targets rather than on one protein recognizing and cutting the entire site alone.
The repair pathway influences whether editing disrupts a gene or introduces a planned sequence change. Non-homologous end joining repairs the double-strand break in a way that can disrupt gene function, whereas homology-directed repair can use the break to introduce a defined sequence change. Selecting or analyzing the resulting repair outcome therefore determines the experiment's genetic consequence.
A basic design begins by selecting the DNA sequence to be modified, then engineering TALE repeats that recognize the chosen target and assembling the corresponding TALEN pair with FokI nuclease domains. After both proteins bind opposite strands and create a break, the cell's repair process generates either gene disruption through non-homologous end joining or a defined change through homology-directed repair.
TALENs are useful when investigators need to examine gene function or create a genetic model with a targeted change. Their applications include disease modeling, production of modified cell lines and organisms, and crop improvement. By directing a break to a chosen sequence, the method links a specific genomic site to an experimentally observable genetic outcome.
In genetics, targeted editing can connect a sequence alteration with changes in gene function, helping researchers study how particular genes contribute to biological traits or disease-related models. The same approach supports engineered cell lines, modified organisms, and crop-related research. Outcomes depend on whether repair disrupts the target gene or introduces a defined sequence change.