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  1. Article ; Online: Livestock breeding for the 21st century

    Chris PROUDFOOT, Gus MCFARLANE, Bruce WHITELAW, Simon LILLICO

    Frontiers of Agricultural Science and Engineering, Vol 7, Iss 2, Pp 129-

    the promise of the editing revolution

    2020  Volume 135

    Abstract: In recent years there has been a veritable explosion in the use of genome editors to create site-specific changes, both in vitro and in vivo, to the genomes of a multitude of species for both basic research and biotechnology. Livestock, which form a ... ...

    Abstract In recent years there has been a veritable explosion in the use of genome editors to create site-specific changes, both in vitro and in vivo, to the genomes of a multitude of species for both basic research and biotechnology. Livestock, which form a vital component of most societies, are no exception. While selective breeding has been hugely successful at enhancing some production traits, the rate of progress is often slow and is limited to variants that exist within the breeding population. Genome editing provides the potential to move traits between breeds, in a single generation, with no impact on existing productivity or to develop de novo phenotypes that tackle intractable issues such as disease. As such, genome editors provide huge potential for ongoing livestock development programs in light of increased demand and disease challenge. This review will highlight some of the more notable agricultural applications of this technology in livestock.
    Keywords cattle|pig|sheep|chicken|aquaculture|crispr ; Agriculture (General) ; S1-972
    Language English
    Publishing date 2020-06-01T00:00:00Z
    Publisher Higher Education Press
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article: Gene targeting, genome editing: from Dolly to editors

    Tan, Wenfang / Chris Proudfoot / Simon G. Lillico / C. Bruce A. Whitelaw

    Transgenic research. 2016 June, v. 25, no. 3

    2016  

    Abstract: One of the most powerful strategies to investigate biology we have as scientists, is the ability to transfer genetic material in a controlled and deliberate manner between organisms. When applied to livestock, applications worthy of commercial venture ... ...

    Abstract One of the most powerful strategies to investigate biology we have as scientists, is the ability to transfer genetic material in a controlled and deliberate manner between organisms. When applied to livestock, applications worthy of commercial venture can be devised. Although initial methods used to generate transgenic livestock resulted in random transgene insertion, the development of SCNT technology enabled homologous recombination gene targeting strategies to be used in livestock. Much has been accomplished using this approach. However, now we have the ability to change a specific base in the genome without leaving any other DNA mark, with no need for a transgene. With the advent of the genome editors this is now possible and like other significant technological leaps, the result is an even greater diversity of possible applications. Indeed, in merely 5 years, these ‘molecular scissors’ have enabled the production of more than 300 differently edited pigs, cattle, sheep and goats. The advent of genome editors has brought genetic engineering of livestock to a position where industry, the public and politicians are all eager to see real use of genetically engineered livestock to address societal needs. Since the first transgenic livestock reported just over three decades ago the field of livestock biotechnology has come a long way—but the most exciting period is just starting.
    Keywords DNA ; biotechnology ; cattle ; gene targeting ; genetic engineering ; goats ; homologous recombination ; sheep ; swine ; transgenes ; transgenic animals
    Language English
    Dates of publication 2016-06
    Size p. 273-287.
    Publishing place Springer International Publishing
    Document type Article
    Note Review
    ZDB-ID 31620-9
    ISSN 1573-9368 ; 0962-8819
    ISSN (online) 1573-9368
    ISSN 0962-8819
    DOI 10.1007/s11248-016-9932-x
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Modelling Neurological Diseases in Large Animals

    Samantha L. Eaton / Fraser Murdoch / Nina M. Rzechorzek / Gerard Thompson / Claudia Hartley / Benjamin Thomas Blacklock / Chris Proudfoot / Simon G. Lillico / Peter Tennant / Adrian Ritchie / James Nixon / Paul M. Brennan / Stefano Guido / Nadia L. Mitchell / David N. Palmer / C. Bruce A. Whitelaw / Jonathan D. Cooper / Thomas M. Wishart

    Cells, Vol 11, Iss 2641, p

    Criteria for Model Selection and Clinical Assessment

    2022  Volume 2641

    Abstract: Issue: The impact of neurological disorders is recognised globally, with one in six people affected in their lifetime and few treatments to slow or halt disease progression. This is due in part to the increasing ageing population, and is confounded by ... ...

    Abstract Issue: The impact of neurological disorders is recognised globally, with one in six people affected in their lifetime and few treatments to slow or halt disease progression. This is due in part to the increasing ageing population, and is confounded by the high failure rate of translation from rodent-derived therapeutics to clinically effective human neurological interventions. Improved translation is demonstrated using higher order mammals with more complex/comparable neuroanatomy. These animals effectually span this translational disparity and increase confidence in factors including routes of administration/dosing and ability to scale, such that potential therapeutics will have successful outcomes when moving to patients. Coupled with advancements in genetic engineering to produce genetically tailored models, livestock are increasingly being used to bridge this translational gap. Approach: In order to aid in standardising characterisation of such models, we provide comprehensive neurological assessment protocols designed to inform on neuroanatomical dysfunction and/or lesion(s) for large animal species. We also describe the applicability of these exams in different large animals to help provide a better understanding of the practicalities of cross species neurological disease modelling. Recommendation: We would encourage the use of these assessments as a reference framework to help standardise neurological clinical scoring of large animal models.
    Keywords neurological disease ; large animal model ; clinical assessment ; model selection criteria ; Biology (General) ; QH301-705.5
    Language English
    Publishing date 2022-08-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Cross-species efficacy of enzyme replacement therapy for CLN1 disease in mice and sheep

    Hemanth R. Nelvagal / Samantha L. Eaton / Sophie H. Wang / Elizabeth M. Eultgen / Keigo Takahashi / Steven Q. Le / Rachel Nesbitt / Joshua T. Dearborn / Nicholas Siano / Ana C. Puhl / Patricia I. Dickson / Gerard Thompson / Fraser Murdoch / Paul M. Brennan / Mark Gray / Stephen N. Greenhalgh / Peter Tennant / Rachael Gregson / Eddie Clutton /
    James Nixon / Chris Proudfoot / Stefano Guido / Simon G. Lillico / C. Bruce A. Whitelaw / Jui-Yun Lu / Sandra L. Hofmann / Sean Ekins / Mark S. Sands / Thomas M. Wishart / Jonathan D. Cooper

    The Journal of Clinical Investigation, Vol 132, Iss

    2022  Volume 20

    Abstract: CLN1 disease, also called infantile neuronal ceroid lipofuscinosis (NCL) or infantile Batten disease, is a fatal neurodegenerative lysosomal storage disorder resulting from mutations in the CLN1 gene encoding the soluble lysosomal enzyme palmitoyl- ... ...

    Abstract CLN1 disease, also called infantile neuronal ceroid lipofuscinosis (NCL) or infantile Batten disease, is a fatal neurodegenerative lysosomal storage disorder resulting from mutations in the CLN1 gene encoding the soluble lysosomal enzyme palmitoyl-protein thioesterase 1 (PPT1). Therapies for CLN1 disease have proven challenging because of the aggressive disease course and the need to treat widespread areas of the brain and spinal cord. Indeed, gene therapy has proven less effective for CLN1 disease than for other similar lysosomal enzyme deficiencies. We therefore tested the efficacy of enzyme replacement therapy (ERT) by administering monthly infusions of recombinant human PPT1 (rhPPT1) to PPT1-deficient mice (Cln1–/–) and CLN1R151X sheep to assess how to potentially scale up for translation. In Cln1–/– mice, intracerebrovascular (i.c.v.) rhPPT1 delivery was the most effective route of administration, resulting in therapeutically relevant CNS levels of PPT1 activity. rhPPT1-treated mice had improved motor function, reduced disease-associated pathology, and diminished neuronal loss. In CLN1R151X sheep, i.c.v. infusions resulted in widespread rhPPT1 distribution and positive treatment effects measured by quantitative structural MRI and neuropathology. This study demonstrates the feasibility and therapeutic efficacy of i.c.v. rhPPT1 ERT. These findings represent a key step toward clinical testing of ERT in children with CLN1 disease and highlight the importance of a cross-species approach to developing a successful treatment strategy.
    Keywords Neuroscience ; Therapeutics ; Medicine ; R
    Language English
    Publishing date 2022-10-01T00:00:00Z
    Publisher American Society for Clinical Investigation
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Substitution of warthog NF-κB motifs into RELA of domestic pigs is not sufficient to confer resilience to African swine fever virus

    Stephen McCleary / Rebecca Strong / Ronan R. McCarthy / Jane C. Edwards / Emma L. Howes / Lisa M. Stevens / Pedro J. Sánchez-Cordón / Alejandro Núñez / Samantha Watson / Alan J. Mileham / Simon G. Lillico / Christine Tait-Burkard / Chris Proudfoot / Maeve Ballantyne / C. Bruce A. Whitelaw / Falko Steinbach / Helen R. Crooke

    Scientific Reports, Vol 10, Iss 1, Pp 1-

    2020  Volume 11

    Abstract: Abstract African swine fever virus (ASFV) causes a lethal, haemorrhagic disease in domestic swine that threatens pig production across the globe. Unlike domestic pigs, warthogs, which are wildlife hosts of the virus, do not succumb to the lethal effects ... ...

    Abstract Abstract African swine fever virus (ASFV) causes a lethal, haemorrhagic disease in domestic swine that threatens pig production across the globe. Unlike domestic pigs, warthogs, which are wildlife hosts of the virus, do not succumb to the lethal effects of infection. There are three amino acid differences between the sequence of the warthog and domestic pig RELA protein; a subunit of the NF-κB transcription factor that plays a key role in regulating the immune response to infections. Domestic pigs with all 3 or 2 of the amino acids from the warthog RELA orthologue have been generated by gene editing. To assess if these variations confer resilience to ASF we established an intranasal challenge model with a moderately virulent ASFV. No difference in clinical, virological or pathological parameters were observed in domestic pigs with the 2 amino acid substitution. Domestic pigs with all 3 amino acids found in warthog RELA were not resilient to ASF but a delay in onset of clinical signs and less viral DNA in blood samples and nasal secretions was observed in some animals. Inclusion of these and additional warthog genetic traits into domestic pigs may be one way to assist in combating the devastating impact of ASFV.
    Keywords Medicine ; R ; Science ; Q
    Subject code 630
    Language English
    Publishing date 2020-06-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Zinc finger recombinases with adaptable DNA sequence specificity.

    Chris Proudfoot / Arlene L McPherson / Andreas F Kolb / W Marshall Stark

    PLoS ONE, Vol 6, Iss 4, p e

    2011  Volume 19537

    Abstract: Site-specific recombinases have become essential tools in genetics and molecular biology for the precise excision or integration of DNA sequences. However, their utility is currently limited to circumstances where the sites recognized by the recombinase ... ...

    Abstract Site-specific recombinases have become essential tools in genetics and molecular biology for the precise excision or integration of DNA sequences. However, their utility is currently limited to circumstances where the sites recognized by the recombinase enzyme have been introduced into the DNA being manipulated, or natural 'pseudosites' are already present. Many new applications would become feasible if recombinase activity could be targeted to chosen sequences in natural genomic DNA. Here we demonstrate efficient site-specific recombination at several sequences taken from a 1.9 kilobasepair locus of biotechnological interest (in the bovine β-casein gene), mediated by zinc finger recombinases (ZFRs), chimaeric enzymes with linked zinc finger (DNA recognition) and recombinase (catalytic) domains. In the "Z-sites" tested here, 22 bp casein gene sequences are flanked by 9 bp motifs recognized by zinc finger domains. Asymmetric Z-sites were recombined by the concomitant action of two ZFRs with different zinc finger DNA-binding specificities, and could be recombined with a heterologous site in the presence of a third recombinase. Our results show that engineered ZFRs may be designed to promote site-specific recombination at many natural DNA sequences.
    Keywords Medicine ; R ; Science ; Q
    Subject code 612
    Language English
    Publishing date 2011-04-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article: Comparison of surrogate reporter systems for enrichment of cells with mutations induced by genome editors

    He, Zuyong / Xuan Shi / Meirui Liu / Guangjie Sun / Chris Proudfoot / C. Bruce A. Whitelaw / Simon G. Lillico / Yaosheng Chen

    Journal of biotechnology. 2016 Mar. 10, v. 221

    2016  

    Abstract: Genome editors are powerful tools that allow modification of the nuclear DNA in eukaryotic cells both in vitro and in vivo. In vitro modified cells are often phenotypically indistinguishable from unmodified cells, hampering their isolation for analysis. ... ...

    Abstract Genome editors are powerful tools that allow modification of the nuclear DNA in eukaryotic cells both in vitro and in vivo. In vitro modified cells are often phenotypically indistinguishable from unmodified cells, hampering their isolation for analysis. Episomal reporters encoding fluorescent proteins can be used for enrichment of modified cells by flow cytometry. Here we compare two surrogate reporters, RGS and SSA, for the enrichment of porcine embryonic fibroblasts containing mutations induced by ZFNs or CRISPR/Cas9. Both systems were effective for enrichment of edited porcine cells with the RGS reporter proving more effective than the SSA reporter. We noted a higher-fold enrichment when editing events were induced by Cas9 compared to those induced by ZFNs, allowing selection at frequencies as high as 70%.
    Keywords eukaryotic cells ; fibroblasts ; flow cytometry ; fluorescent proteins ; mutagenesis ; nuclear genome ; swine
    Language English
    Dates of publication 2016-0310
    Size p. 49-54.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 843647-2
    ISSN 1873-4863 ; 0168-1656 ; 1389-0352
    ISSN (online) 1873-4863
    ISSN 0168-1656 ; 1389-0352
    DOI 10.1016/j.jbiotec.2016.01.009
    Database NAL-Catalogue (AGRICOLA)

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