32
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Engineering large animal models of human disease

      review-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          The recent development of gene editing tools and methodology for use in livestock enables the production of new animal disease models. These tools facilitate site‐specific mutation of the genome, allowing animals carrying known human disease mutations to be produced. In this review, we describe the various gene editing tools and how they can be used for a range of large animal models of diseases. This genomic technology is in its infancy but the expectation is that through the use of gene editing tools we will see a dramatic increase in animal model resources available for both the study of human disease and the translation of this knowledge into the clinic. Comparative pathology will be central to the productive use of these animal models and the successful translation of new therapeutic strategies. © 2015 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

          Related collections

          Most cited references85

          • Record: found
          • Abstract: found
          • Article: found
          Is Open Access

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

          TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. We present a method and reagents for efficiently assembling TALEN constructs with custom repeat arrays. We also describe design guidelines based on naturally occurring TAL effectors and their binding sites. Using software that applies these guidelines, in nine genes from plants, animals and protists, we found candidate cleavage sites on average every 35 bp. Each of 15 sites selected from this set was cleaved in a yeast-based assay with TALEN pairs constructed with our reagents. We used two of the TALEN pairs to mutate HPRT1 in human cells and ADH1 in Arabidopsis thaliana protoplasts. Our reagents include a plasmid construct for making custom TAL effectors and one for TAL effector fusions to additional proteins of interest. Using the former, we constructed de novo a functional analog of AvrHah1 of Xanthomonas gardneri. The complete plasmid set is available through the non-profit repository AddGene and a web-based version of our software is freely accessible online.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            A TALE nuclease architecture for efficient genome editing.

            Nucleases that cleave unique genomic sequences in living cells can be used for targeted gene editing and mutagenesis. Here we develop a strategy for generating such reagents based on transcription activator-like effector (TALE) proteins from Xanthomonas. 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 of designed TALE transcription factors and nucleases for the targeted regulation and modification of endogenous genes.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              FLASH Assembly of TALENs Enables High-Throughput Genome Editing

              Engineered transcription activator-like effector nucleases (TALENs) have shown promise as facile and broadly applicable genome editing tools. However, no publicly available high-throughput method for constructing TALENs has been published and large-scale assessments of the success rate and targeting range of the technology remain lacking. Here we describe the Fast Ligation-based Automatable Solid-phase High-throughput (FLASH) platform, a rapid and cost-effective method we developed to enable large-scale assembly of TALENs. We tested 48 FLASH-assembled TALEN pairs in a human cell-based EGFP reporter system and found that all 48 possessed efficient gene modification activities. We also used FLASH to assemble TALENs for 96 endogenous human genes implicated in cancer and/or epigenetic regulation and found that 84 pairs were able to efficiently introduce targeted alterations. Our results establish the robustness of TALEN technology and demonstrate that FLASH facilitates high-throughput genome editing at a scale not currently possible with engineered zinc-finger nucleases or meganucleases.
                Bookmark

                Author and article information

                Journal
                J Pathol
                J. Pathol
                10.1002/(ISSN)1096-9896
                PATH
                The Journal of Pathology
                John Wiley & Sons, Ltd (Chichester, UK )
                0022-3417
                1096-9896
                28 November 2015
                January 2016
                : 238
                : 2 , Models of Human Disease ( doiID: 10.1002/path.2016.238.issue-2 )
                : 247-256
                Affiliations
                [ 1 ] The Roslin Institute and Royal (Dick) School of Veterinary Science, Easter Bush CampusUniversity of Edinburgh Edinburgh EH25 9RGUK
                [ 2 ]Animal Bioscience and Biotechnology Laboratory, ARS Beltsville MD 20705USA
                [ 3 ]Department of Animal and Avian Sciences Beltsville MD 20742USA
                Author notes
                [*] [* ]Correspondence to: B Whitelaw, The Roslin Institute and Royal (Dick) School of Veterinary Science, Easter Bush Campus, University of Edinburgh, Edinburgh, EH25 9RG, UK. E‐mail: bruce.whitelaw@ 123456roslin.ed.ac.uk Or B Telugu, 2121 ANSC Building, University of Maryland, College Park, MD 20742, USA. E‐mail: btelugu@ 123456umd.edu
                Article
                PATH4648 15-583.R1
                10.1002/path.4648
                4737318
                26414877
                a16af529-574f-4fc9-acfb-df6c05645db7
                © 2015 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 07 August 2015
                : 15 September 2015
                : 22 September 2015
                Page count
                Pages: 10
                Funding
                Funded by: BBSRC ISPG
                Funded by: BBSRC IPA
                Award ID: BB/L007371/1
                Funded by: Genus plc
                Funded by: NIFA Dual Purpose with Dual Benefit Grant
                Award ID: 2015‐67015‐22845
                Categories
                Invited Review
                Invited Reviews
                Custom metadata
                2.0
                path4648
                January 2016
                Converter:WILEY_ML3GV2_TO_NLMPMC version:4.7.5 mode:remove_FC converted:28.01.2016

                Pathology
                crispr,gene editing,livestock,pathology,pigs,scnt,talen,zfn,zygote
                Pathology
                crispr, gene editing, livestock, pathology, pigs, scnt, talen, zfn, zygote

                Comments

                Comment on this article