A TALE nuclease architecture for efficient genome editing

  title={A TALE nuclease architecture for efficient genome editing},
  author={Jeffrey C. Miller and Siyuan Tan and Guijuan Qiao and Kyle A. Barlow and Jianbin Wang and Danny F Xia and Xiangdong Meng and David E. Paschon and Elo Leung and Sarah Hinkley and Gladys P. Dulay and Kevin L Hua and Irina Ankoudinova and Gregory J Cost and Fyodor D. Urnov and H. Steve Zhang and Michael C. Holmes and Lei Zhang and Philip D. Gregory and Edward J Rebar},
  journal={Nature Biotechnology},
Nucleases that cleave unique genomic sequences in living cells can be used for targeted gene editing and mutagenesis. [] Key Result We identify TALE truncation variants that efficiently cleave DNA when linked to the catalytic domain of FokI and use these nucleases to generate discrete edits or small deletions within endogenous human NTF3 and CCR5 genes at efficiencies of up to 25%. We further show that designed TALEs can regulate endogenous mammalian genes. These studies demonstrate the effective application…

TALEN-Mediated Mutagenesis and Genome Editing.

A detailed in silico design tool for zebrafish genome editing for TALENs and CRISPR/Cas9 custom restriction enzymes using Mojo Hand 2.0 software is described.

TALENs: a widely applicable technology for targeted genome editing

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.

A novel TALE nuclease scaffold enables high genome editing activity in combination with low toxicity

The combination of high nuclease activity with reduced cytotoxicity and the simple design process marks TALENs as a key technology platform for targeted modifications of complex genomes.

Genome-Editing Technologies: Principles and Applications.

Three foundational technologies-clustered regularly interspaced short palindromic repeats (CRISPR)- CRISPR-associated protein 9 (Cas9), transcription activator-like effector nucleases (TALENs), and zinc-finger nuclease (ZFNs) are reviewed, illustrating how this technology can complement targeted nucleases for synthetic biology and gene therapy.

[TALE nuclease engineering and targeted genome modification].

The recent progresses and prospects of TALEN technology are summarized, with an emphasis on its structure, function, and construction strategies, as well as a collection of species and genes that have been successfully modified by TALens, especially the application in zebrafish.

Genome editing comes of age

This work highlights the key advances that set the foundation for the rapid and widespread implementation of CRISPR–Cas9 genome editing approaches that has revolutionized the field.

Design, evaluation, and screening methods for efficient targeted mutagenesis with transcription activator‐like effector nucleases in medaka

It is found that a specific pattern of mutations is dominant for TALENs harboring several base pairs of homologous sequences in target sequence, and a 5′ T, upstream of each TALen‐binding sequence, is not essential for genomic DNA cleavage.



Critical parameters for genome editing using zinc finger nucleases.

This review will discuss the advances in engineering custom zinc-finger nucleases and their application in stimulating homologous recombination in higher eukaryotic cells at efficiencies approaching 1 in 2 cells.

Targeted genome editing in human cells with zinc finger nucleases constructed via modular assembly.

This work synthesized and tested hundreds of ZFNs to target dozens of different sites in the human CCR5 gene-a co-receptor required for HIV infection-and found that many of these nucleases induced site-specific mutations in the C CR5 sequence.

An improved zinc-finger nuclease architecture for highly specific genome editing

Using structure-based design, two variant ZFNs are engineer that modify a native endogenous locus as efficiently as the parental architecture, but with a >40-fold reduction in homodimer function and much lower levels of genome-wide cleavage.

Genome editing with engineered zinc finger nucleases

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.

Targeting DNA Double-Strand Breaks with TAL Effector Nucleases

A new class of sequence-specific nucleases created by fusing transcription activator-like effectors (TALEs) to the catalytic domain of the FokI endonuclease is reported.

Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures

The development and application of a yeast-based selection system designed to functionally interrogate the ZFN dimer interface is reported, identified critical residues involved in dimerization through the isolation of cold-sensitive nuclease domains, and used to engineer ZFNs that have superior cleavage activity while suppressing homodimerization.

Zinc Finger Nucleases as tools to understand and treat human diseases

This review discusses the development and use of designer zinc finger proteins (ZFPs) as sequence specific tools, and the utility of other functional domains, such as transcriptional activators and repressors, and highlights how these are being used as discovery and therapeutic tools.

TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain

The creation and initial characterization of a group of rare-cutting, site-specific DNA nucleases produced by fusion of the restriction enzyme FokI endonuclease domain (FN) with the high-specificity DNA-binding domains of AvrXa7 and PthXo1 are reported.

Zinc-finger nucleases: the next generation emerges.

  • T. CathomenJ. Joung
  • Biology
    Molecular therapy : the journal of the American Society of Gene Therapy
  • 2008
Recent advances in gene targeting, and upcoming challenges for, this emerging technology are reviewed and future experimental work that will be needed to bring ZFNs safely into a clinical setting are discussed.

A rapid and general assay for monitoring endogenous gene modification.

A simple procedure for quantifying mutations that result from DNA double-strand break repair via non-homologous end joining based on the ability of the Surveyor nuclease to selectively cleave distorted duplex DNA formed via cross-annealing of mutated and wild-type sequence is described.