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Open AccessJournal ArticleDOI

TALENs: a widely applicable technology for targeted genome editing

J. Keith Joung, +1 more
- 01 Jan 2013 - 
- Vol. 14, Iss: 1, pp 49-55
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TLDR
The newly-developed transcription activator-like effector nucleases (TALENs) comprise a nonspecific DNA-cleaving nuclease fused to a DNA-binding domain that can be easily engineered so that TALens can target essentially any sequence.
Abstract
Engineered nucleases enable the targeted alteration of nearly any gene in a wide range of cell types and organisms. The newly-developed transcription activator-like effector nucleases (TALENs) comprise a nonspecific DNA-cleaving nuclease fused to a DNA-binding domain that can be easily engineered so that TALENs can target essentially any sequence. The capability to quickly and efficiently alter genes using TALENs promises to have profound impacts on biological research and to yield potential therapeutic strategies for genetic diseases.

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Citations
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Conditional knockouts generated by engineered CRISPR-Cas9 endonuclease reveal the roles of coronin in C. elegans neural development.

TL;DR: An essential embryonic gene, Coronin, which is associated with human neurobehavioral dysfunction, regulates actin organization and cell morphology during C. elegans postembryonic neuroblast migration and neuritogenesis is shown.
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Genome-wide identification of CRISPR/Cas9 off-targets in human genome

TL;DR: It is demonstrated that CRISPR/Cas9 had crRNA-specific off-target binding activities in human genome, and hypothesized that the Cas9/crRNA complex must first bind significantly to those off-targets, which could be revealed by chromatin immunoprecipitation and high throughput genome sequencing (ChIP-seq).
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Engineering the AAVS1 locus for consistent and scalable transgene expression in human iPSCs and their differentiated derivatives.

TL;DR: The AAVS1 targeting protocol is outlined, using standardized donor vectors and construction methods, as well as practical considerations for iPSC culture, drug selection, and genotyping, to introduce transgenes into pre-defined loci and overcome random position effects.
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DNA Methylation in Social Insects: How Epigenetics Can Control Behavior and Longevity

TL;DR: Recent advances in DNA methylation research are summarized and its significance in understanding of the epigenetic underpinnings of behavior and longevity is discussed.
Book ChapterDOI

Biology and Diseases of Mice

TL;DR: Today’s laboratory mouse, Mus musculus, has its origins as the ‘house mouse’ of North America and Europe and since the mid-1980s, additional strains have been developed from Asian mice.
References
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Journal ArticleDOI

Breaking the Code of DNA Binding Specificity of TAL-Type III Effectors

TL;DR: The functionality of a distinct type of DNA binding domain is described and allows the design ofDNA binding domains for biotechnology.
Journal ArticleDOI

Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting

TL;DR: A method and reagents for efficiently assembling TALEN constructs with custom repeat arrays are presented and design guidelines based on naturally occurring TAL effectors and their binding sites are described.
Journal ArticleDOI

A TALE nuclease architecture for efficient genome editing

TL;DR: This study identifies TALE truncation variants that efficiently cleave DNA when linked to the catalytic domain of FokI and uses them to generate discrete edits or small deletions within endogenous human NTF3 and CCR5 genes at efficiencies of up to 25%.
Journal ArticleDOI

Genome editing with engineered zinc finger nucleases

TL;DR: A broad range of outcomes has resulted from the application of the same core technology: targeted genome cleavage by engineered, sequence-specific zinc finger nucleases followed by gene modification during subsequent repair.
Journal ArticleDOI

A Simple Cipher Governs DNA Recognition by TAL Effectors

TL;DR: It is shown that a repeat-variable pair of residues specifies the nucleotides in the target site, one pair to one nucleotide, with no apparent context dependence, which represents a previously unknown mechanism for protein-DNA recognition that explains TAL effector specificity, enables target site prediction, and opens prospects for use of TAL effects in research and biotechnology.
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