E-TALEN: a web tool to design TALENs for genome engineering

Heigwer, Florian; Kerr, Gráinne; Walther, Nike; Glaeser, Kathrin; Pelz, Oliver; Breinig, Marco; Boutros, Michael

doi:10.1093/nar/gkt789

Cited by 60 publications

(22 citation statements)

References 22 publications

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“…E-TALEN allows a choice among several assembly kits to inform design, and both tools allow the user to specify the end purpose of the experiment (knock-out, N- or C-terminal tagging) as well as other tool-specific criteria [75]. …”

Section: Designing and Engineering Talens And Crispr Reagentsmentioning

confidence: 99%

Targeted genome engineering techniques in Drosophila

Beumer

Carroll

2014

Methods

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For a century, Drosophila has been a favored organism for genetic research. However, the array of materials and methods available to the Drosophila worker has expanded dramatically in the last decade. The most common gene targeting tools, zinc finger nucleases, TALENs, and RNA-guided CRISPR/Cas9, have all been adapted for use in Drosophila, both for simple mutagenesis and for gene editing via homologous recombination. For each tool, there exist a number of web sites, design applications, and delivery methods. The successful application of any of these tools also requires an understanding of methods for detecting successful genome modifications. This article provides an overview of the available gene targeting tools and their application in Drosophila. In lieu of simply providing a protocol for gene targeting, we direct the researcher to resources that will allow access to the latest research in this rapidly evolving field.

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Section: Designing and Engineering Talens And Crispr Reagentsmentioning

confidence: 99%

Targeted genome engineering techniques in Drosophila

Beumer

Carroll

2014

Methods

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“…Use your favorite TALE design software [For example E-TALEN (Heigwer et al, 2013) or MojoHand (Neff et al, 2013)] to select the precise location of TALE binding within the target region. If the goal is to generate frameshift mutations in an ORF, then attention should be paid to several other factors such as the presence of alternative start codons, alternative promoters or splice variants which may confound experimental outcomes.…”

Section: Methodsmentioning

confidence: 99%

Targeted genome editing in Aedes aegypti using TALENs

Aryan

Myles

Adelman

2014

Methods

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The Culicine mosquito, Aedes aegypti, is both a major vector of arthropod-borne viruses (arboviruses) and a genetic model organism for arbovirus transmission. TALE nucleases (TALENs), a group of artificial enzymes capable of generating site-specific DNA lesions, consist of a non-specific FokI endonuclease cleavage domain fused to an engineered DNA binding domain specific to a target site. While TALENs have become an important tool for targeted gene disruption in a variety of organisms, application to the mosquito genome is a new approach. We recently described the use of TALENs to perform heritable genetic disruptions in Ae. aegypti. Here, we provide detailed methods that will allow other research laboratories to capitalize on the potential of this technology for understanding mosquito gene function. We describe target site selection, transient embryo-based assays to rapidly assess TALEN activity, embryonic microinjection and downstream screening steps to identify target site mutations.

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“…There are two approaches to reduce off-target editing: 1) increasing the specificity of each TALENs DNA binding domain; and 2) avoiding off-target sites found elsewhere in the human genome. In target site selection, in silico tools like Tal Effector Nucleotide Targeter 2.0, Prognos, CHOPCHOP, E-TALEN, TALEN-NT, TALENoffer, and SAPTA predict potential off-target sites with good accuracy (Doyle et al 2012; Grau et al 2013; Heigwer et al 2013; Fine et al 2014; Lin et al 2014; Montague et al 2014). …”

Section: Host and Viral Targets For Gene Editingmentioning

confidence: 99%

TALEN gene editing takes aim on HIV

et al. 2016

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Transcription activator-like effector nucleases (TALENs) are one of several types of programmable, engineered nucleases that bind and cleave specific DNA sequences. Cellular machinery repairs the cleaved DNA by introducing indels. In this review, we emphasize the potential, explore progress, and identify challenges in using TALENs as a therapeutic tool to treat HIV infection. TALENs have less off-target editing and can be more effective at tolerating HIV escape mutations than CRISPR/Cas-9. Scientists have explored TALEN-mediated editing of host genes such as viral entry receptors (CCR5 and CXCR4), and a protein involved in proviral integration (LEDGF/p75). Viral targets include the proviral DNA, particularly, and the long terminal repeats. Major challenges with translating gene therapy from bench to bedside are improving cleavage efficiency and delivery, while minimizing off-target editing, cytotoxicity, and immunogenicity. However, rapid improvements in TALEN technology are enhancing cleavage efficiency and specificity. Therapeutic testing in animal models of HIV infection will help determine whether TALENs are a viable HIV treatment therapy. TALENs or other engineered nucleases could shift the therapeutic paradigm from life-long antiretroviral therapy towards eradication of HIV infection.

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E-TALEN: a web tool to design TALENs for genome engineering

Cited by 60 publications

References 22 publications

Targeted genome engineering techniques in Drosophila

Targeted genome engineering techniques in Drosophila

Targeted genome editing in Aedes aegypti using TALENs

TALEN gene editing takes aim on HIV

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