TALENs: a widely applicable technology for targeted genome editing
J. Keith Joung,Jeffry D. Sander +1 more
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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.read more
Citations
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CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes
Luke A. Gilbert,Matthew H. Larson,Leonardo Morsut,Zairan Liu,Gloria A. Brar,Sandra Elizabeth Torres,Noam Stern-Ginossar,Onn Brandman,Evan H. Whitehead,Jennifer A. Doudna,Wendell A. Lim,Jonathan S. Weissman,Lei S. Qi +12 more
TL;DR: The results establish that the CRISPR system can be used as a modular and flexible DNA-binding platform for the recruitment of proteins to a target DNA sequence, revealing the potential of CRISpri as a general tool for the precise regulation of gene expression in eukaryotic cells.
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CRISPR-Cas systems for editing, regulating and targeting genomes
Jeffry D. Sander,J. Keith Joung +1 more
TL;DR: A modified version of the CRISPR-Cas9 system has been developed to recruit heterologous domains that can regulate endogenous gene expression or label specific genomic loci in living cells, which will undoubtedly transform biological research and spur the development of novel molecular therapeutics for human disease.
Journal ArticleDOI
Efficient genome editing in zebrafish using a CRISPR-Cas system
Woong Y. Hwang,Yanfang Fu,Deepak Reyon,Morgan L. Maeder,Shengdar Q. Tsai,Jeffry D. Sander,Randall T. Peterson,Randall T. Peterson,Jing-Ruey J. Yeh,J. Keith Joung +9 more
TL;DR: It is shown that the CRISPR-Cas system functions in vivo to induce targeted genetic modifications in zebrafish embryos with efficiencies similar to those obtained using zinc finger nucleases and transcription activator-like effector nucleases.
Journal ArticleDOI
Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system
TL;DR: An improved CRISPR/Cas system in zebra fish with custom guide RNAs and a zebrafish codon-optimized Cas9 protein that efficiently targeted a reporter transgene Tg(-5.1mnx1:egfp) and four endogenous loci and five genomic loci, resulting in multiple loss-of-function phenotypes in the same injected fish.
Journal ArticleDOI
A guide to genome engineering with programmable nucleases
Hyongbum Kim,Jin-Soo Kim +1 more
TL;DR: Known nuclease-specific features are essential for researchers to choose the most appropriate tool for a range of applications, including their composition, targetable sites, specificities and mutation signatures, among other characteristics.
References
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Journal ArticleDOI
Improved somatic mutagenesis in zebrafish using transcription activator-like effector nucleases (TALENs).
Finola E. Moore,Deepak Reyon,Jeffry D. Sander,Sarah Martinez,Jessica S. Blackburn,Cyd Khayter,Cherie L. Ramirez,J. Keith Joung,David M. Langenau +8 more
TL;DR: The results suggest that TALEN technology provides a robust alternative to CoDA ZFNs for inducing targeted gene-inactivation in zebrafish, making it a preferred technology for creating targeted knockout mutants inZebrafish.
Journal ArticleDOI
Designing Transcription Factor Architectures for Drug Discovery
TL;DR: The potential of designer transcription factors to alter the transcription of specific genes, to discover new genes, and to induce phenotypes in cells and organisms is now being applied in the areas of molecular therapeutics, pharmacology, biotechnology, and functional genomics.
Journal ArticleDOI
Targeted chromosomal duplications and inversions in the human genome using zinc finger nucleases.
TL;DR: It is found that zinc finger nucleases designed to target two different sites in a human chromosome could introduce two concurrent double-strand breaks, whose repair via non-homologous end-joining (NHEJ) gives rise to targeted duplications and inversions of the genomic segments of up to a mega base pair in length between the two sites.
Journal ArticleDOI
Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.
Tatiana Flisikowska,Irmgard Thorey,Sonja Offner,Francesca Ros,Valeria Lifke,Bryan Zeitler,Oswald Rottmann,Anna I Vincent,Lei Zhang,Shirin S Jenkins,Helmut Niersbach,Alexander Kind,Philip D. Gregory,Angelika Schnieke,Josef Platzer +14 more
TL;DR: It is demonstrated that zinc finger nucleases introduced into fertilized oocytes can inactivate a chosen gene by mutagenesis and also mediate precise homologous recombination with a DNA gene-targeting vector to achieve the first gene knockout and targeted sequence replacement in rabbits.
Journal ArticleDOI
Synergy between adjacent zinc fingers in sequence-specific DNA recognition
Mark Isalan,Yen Choo,Aaron Klug +2 more
TL;DR: It is shown that Zif268-like zinc fingers can specify overlapping 4-bp subsites, and that sequence specificity at the boundary between subsites arises from synergy between adjacent fingers.
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