title: The genome editing revolution: A CRISPR-Cas TALE off-target story.
abstract: In the last 10 years, we have witnessed a blooming of targeted genome editing systems and applications. The area was revolutionized by the discovery and characterization of the transcription activator-like effector proteins, which are easier to engineer to target new DNA sequences than the previously available DNA binding templates, zinc fingers and meganucleases. Recently, the area experimented a quantum leap because of the introduction of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system (clustered regularly interspaced short palindromic sequence). This ribonucleoprotein complex protects bacteria from invading DNAs, and it was adapted to be used in genome editing. The CRISPR ribonucleic acid (RNA) molecule guides to the specific DNA site the Cas9 nuclease to cleave the DNA target. Two years and more than 1000 publications later, the CRISPR-Cas system has become the main tool for genome editing in many laboratories. Currently the targeted genome editing technology has been used in many fields and may be a possible approach for human gene therapy. Furthermore, it can also be used to modifying the genomes of model organisms for studying human pathways or to improve key organisms for biotechnological applications, such as plants, livestock genome as well as yeasts and bacterial strains.
authors.affiliation: Novo Nordisk Foundation Center for Protein Research, Protein Structure and Function Programme, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Stefano
authors.initials: S
authors.lastname: Stella
authors.affiliation: Novo Nordisk Foundation Center for Protein Research, Protein Structure and Function Programme, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Montoya
mesh_terms: D000818:Animals; D001419:Bacteria; D064113:CRISPR-Cas Systems; D056890:Eukaryota; D000072669:Gene Editing / Q000379:methods*; D006801:Humans
keywords: CRISPR-Cas9; TALE; engineered nucleases; genome editing; specificity