ZFN, TALEN and CRISPR/Cas-based methods for genome engineering

By T. Gaj, C. Gersbach, Carlos F. Barbas III. Trends in Biotechnology, 2013

3,618 citations recorded by the source corpus.

Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) comprise a powerful class of tools that are redefining the boundaries of biological research. These chimeric nucleases are composed of programmable, sequence-specific DNA-binding modules linked to a non-specific DNA cleavage domain. ZFNs and TALENs enable a broad range of genetic modifications by inducing DNA double-strand breaks that stimulate error-prone non-homologous end joining or homology-directed repair at specific genomic locations. Here, we review achievements made possible by site-specific nuclease technologies and discuss applications of these reagents for genetic analysis and manipulation. In addition, we highlight the therapeutic potential of ZFNs and TALENs and discuss future prospects for the field, including the emergence of CRISPR/Cas-based RNA-guided DNA endonucleases.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/59349f554fcc856446afb7ba943bcfc1d8928f1b

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Citations in the corpus

Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System (2013, neutral): “Further reduction of the required number of sgRNAs could be implemented by decreasing the background level of dCas9-EGFP or improving the imaging sensitivity (Gaj et al., 2013).”
Biology and Diseases of Mice (2015, positive): “Recent advances in engineered endonuclease (EE) technology, including zinc finger nucleases (ZFNs), LABORATORY ANIMAL MEDICINE transcription activator-like effector nucleases (TALENs), and RNA-guided endonucleases (RGENs), have revolutionized the field of transgenics (Sung et al., 2014; Wijshake et ”
Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair (2014, background): “Methods of genome engineering are becoming increasingly powerful owing to breakthroughs in the design of artificial nucleases that induce site-specific double-strand breaks (DSBs) in the genome (Gaj et al. 2013).”
Easy quantitative assessment of genome editing by sequence trace decomposition (2014, methodology): “(b) Overview of TIDE algorithm and output, which consists of three main steps: (1) Visualization of aberrant sequence signal in control (black) and treated sample (green), the expected break site (vertical dotted line) and the region used for decomposition (gray bar); (2) Decomposition yielding the ”
mRNA-based therapeutics: powerful and versatile tools to combat diseases (2022, background): “mRNA drugs modulate cellular genomic information by encoding artificial endonucleases, such as ZFNs, TALENs, and more recently CRISPR/Cas nuclease systems.(462) Generally, the three mRNA-encoded endonucleases were designed to achieve insertions/deletions (indels) and mutations by introducing a targe”
Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice (2013, background): “Within the past 3 years, TAL effector nuclease (TALEN) technology has come to the fore as a more tractable system for targeted gene disruption and, in some cases, gene replacement [reviewed in (13,14)].”