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  • 1
    Online Resource
    Online Resource
    Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep
    Springer Science and Business Media LLC ; 2016
    In:  Scientific Reports Vol. 6, No. 1 ( 2016-08-26)
    Details
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 6, No. 1 ( 2016-08-26)
    Abstract: The CRISPR/Cas9 system provides a flexible approach for genome engineering of genetic loci. Here, we successfully achieved precise gene targeting in sheep by co-injecting one-cell-stage embryos with Cas9 mRNA and RNA guides targeting three genes ( MSTN , ASIP and BCO2 ). We carefully examined the sgRNAs:Cas9-mediated targeting effects in injected embryos, somatic tissues, as well as gonads via cloning and sequencing. The targeting efficiencies in these three genes were within the range of 27–33% in generated lambs and that of simultaneously targeting the three genes was 5.6%, which demonstrated that micro-injection of zygotes is an efficient approach for generating gene-modified sheep. Interestingly, we observed that disruption of the MSTN gene resulted in the desired muscle hypertrophy that is characterized by enlarged myofibers, thereby providing the first detailed evidence supporting that gene modifications had occurred at both the genetic and morphological levels. In addition, prescreening for the off-target effect of sgRNAs was performed on fibroblasts before microinjection, to ensure that no detectable off-target mutations from founder animals existed. Our findings suggested that the CRISPR/Cas9 method can be exploited as a powerful tool for livestock improvement by simultaneously targeting multiple genes that are responsible for economically significant traits.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    URL: Article
    DOI: 10.1038/srep32271
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2016
    detail.hit.zdb_id: 2615211-3
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  • 2
    Online Resource
    Online Resource
    Efficient generation of goats with defined point mutation (I397V) in GDF9 through CRISPR/Cas9
    CSIRO Publishing ; 2018
    In:  Reproduction, Fertility and Development Vol. 30, No. 2 ( 2018), p. 307-
    Details
    In: Reproduction, Fertility and Development, CSIRO Publishing, Vol. 30, No. 2 ( 2018), p. 307-
    Abstract: The recent emergence of the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system has attracted significant attention for its potential to improve traits of agricultural importance. However, most applications in livestock species to date have depended on aberrant DNA repair to generate frameshifting indels. Whether this genomic engineering technique involving homology-dependent repair (HDR) can be used to introduce defined point mutations has been less explored. Previously, we reported a G→A point mutation (g.231A 〉 G, p.Val397Ile) in the growth differentiation factor 9 (GDF9) gene that has a large effect on the litter size of cashmere goats. In the present study we report that by co-injecting synthesised RNAs and single-stranded oligo deoxynucleotide (ssODN) donor sequences into goat zygotes, we successfully introduced defined point mutations resulting in single amino acid substitutions in the proteins as expected. The efficiency of this precise single-nucleotide substitution in newborn kids was as high as 24% (4/17), indicating that ssODN-directed HDR via zygote injection is efficient at introducing point mutations in the goat genome. The findings of the present study further highlight the complex genome modifications facilitated by the CRISPR/Cas9 system, which is able to introduce defined point mutations. This represents a significant development for the improvement of reproduction traits in goats, as well as for validating the roles of specific nucleotides in functional genetic elements in large animals.
    Type of Medium: Online Resource
    ISSN: 1031-3613
    URL: Article
    DOI: 10.1071/RD17068
    Language: English
    Publisher: CSIRO Publishing
    Publication Date: 2018
    SSG: 12
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  • 3
    Online Resource
    Online Resource
    Base pair editing in goat: nonsense codon introgression into FGF 5 results in longer hair
    Wiley ; 2019
    In:  The FEBS Journal Vol. 286, No. 23 ( 2019-12), p. 4675-4692
    Details
    In: The FEBS Journal, Wiley, Vol. 286, No. 23 ( 2019-12), p. 4675-4692
    Abstract: The ability to alter single bases without homology directed repair ( HDR ) of double‐strand breaks provides a potential solution for editing livestock genomes for economic traits, which are often multigenic. Progress toward multiplex editing in large animals has been hampered by the costly inefficiencies of HDR via microinjection of in vitro manipulated embryos. Here, we designed sg RNA s to induce nonsense codons (C‐to‐T transitions) at four target sites in caprine FGF 5 , which is a crucial regulator of hair length in mammals. Initial transfections of the third generation Base Editor ( BE 3) plasmid and four different sg RNA s into caprine fibroblasts were ineffective in altering FGF 5 . In contrast, all five progenies produced from microinjected single‐cell embryos had alleles with a targeted nonsense mutation. The effectiveness of BE 3 to make single base changes varied considerably based on sg RNA design. In addition, the rate of mosaicism differed between animals, target sites, and tissue type. The phenotypic effects on hair fiber were characterized by hematoxylin and eosin, immunofluorescence staining, and western blotting. Differences in morphology were detectable, even though mosaicism was probably affecting the levels of FGF 5 expression. PCR amplicon and whole‐genome resequencing analyses for off‐target changes caused by BE 3 were low at a genome‐wide scale. This study provided the first evidence of base editing in large mammals produced from microinjected single‐cell embryos. Our results support further optimization of BE s for introgressing complex human disease alleles into large animal models, to evaluate potential genetic improvement of complex health and production traits in a single generation.
    Type of Medium: Online Resource
    ISSN: 1742-464X , 1742-4658
    URL: Issue
    URL: Article
    DOI: 10.1111/febs.v286.23
    DOI: 10.1111/febs.14983
    Language: English
    Publisher: Wiley
    Publication Date: 2019
    detail.hit.zdb_id: 2172518-4
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Markhor-derived Introgression of a Genomic Region Encompassing PAPSS2 Confers High-altitude Adaptability in Tibetan Goats
    Oxford University Press (OUP) ; 2022
    In:  Molecular Biology and Evolution Vol. 39, No. 12 ( 2022-12-05)
    Details
    In: Molecular Biology and Evolution, Oxford University Press (OUP), Vol. 39, No. 12 ( 2022-12-05)
    Abstract: Understanding the genetic mechanism of how animals adapt to extreme conditions is fundamental to determine the relationship between molecular evolution and changing environments. Goat is one of the first domesticated species and has evolved rapidly to adapt to diverse environments, including harsh high-altitude conditions with low temperature and poor oxygen supply but strong ultraviolet radiation. Here, we analyzed 331 genomes of domestic goats and wild caprid species living at varying altitudes (high & gt; 3000 m above sea level and low & lt; 1200 m), along with a reference-guided chromosome-scale assembly (contig-N50: 90.4 Mb) of a female Tibetan goat genome based on PacBio HiFi long reads, to dissect the genetic determinants underlying their adaptation to harsh conditions on the Qinghai-Tibetan Plateau (QTP). Population genomic analyses combined with genome-wide association studies (GWAS) revealed a genomic region harboring the 3′-phosphoadenosine 5′-phosphosulfate synthase 2 (PAPSS2) gene showing strong association with high-altitude adaptability (PGWAS = 3.62 × 10−25) in Tibetan goats. Transcriptomic data from 13 tissues revealed that PAPSS2 was implicated in hypoxia-related pathways in Tibetan goats. We further verified potential functional role of PAPSS2 in response to hypoxia in PAPSS2-deficient cells. Introgression analyses suggested that the PAPSS2 haplotype conferring the high-altitude adaptability in Tibetan goats originated from a recent hybridization between goats and a wild caprid species, the markhor (Capra falconeri). In conclusion, our results uncover a hitherto unknown contribution of PAPSS2 to high-altitude adaptability in Tibetan goats on QTP, following interspecific introgression and natural selection.
    Type of Medium: Online Resource
    ISSN: 0737-4038 , 1537-1719
    URL: Article
    DOI: 10.1093/molbev/msac253
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2022
    detail.hit.zdb_id: 2024221-9
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    A Mixed-Integer Linear Programming Model for the Station Capacity Allocation Problem of a Star-Tree Pipe Network
    American Society of Civil Engineers (ASCE) ; 2023
    In:  Journal of Pipeline Systems Engineering and Practice Vol. 14, No. 1 ( 2023-02)
    Details
    In: Journal of Pipeline Systems Engineering and Practice, American Society of Civil Engineers (ASCE), Vol. 14, No. 1 ( 2023-02)
    Type of Medium: Online Resource
    ISSN: 1949-1190 , 1949-1204
    URL: Article
    DOI: 10.1061/(ASCE)PS.1949-1204.0000692
    Language: English
    Publisher: American Society of Civil Engineers (ASCE)
    Publication Date: 2023
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  • 6
    Online Resource
    Online Resource
    Inactivation of the MSTN gene expression changes the composition and function of the gut microbiome in sheep
    Springer Science and Business Media LLC ; 2022
    In:  BMC Microbiology Vol. 22, No. 1 ( 2022-11-11)
    Details
    In: BMC Microbiology, Springer Science and Business Media LLC, Vol. 22, No. 1 ( 2022-11-11)
    Abstract: Myostatin ( MSTN ) negatively regulates the muscle growth in animals and MSTN deficient sheep have been widely reported previously. The goal of this study was to explore how MSTN inactivation influences their gut microbiota composition and potential functions. Results We compared the slaughter parameters and meat quality of 3 MSTN -edited male sheep and 3 wild-type male sheep, and analyzed the gut microbiome of the MSTN -edited sheep (8 female and 8 male sheep) and wild-type sheep (8 female and 8 male sheep) through metagenomic sequencing. The results showed that the body weight, carcass weight and eye muscle area of MSTN -edited sheep were significantly higher, but there were no significant differences in the meat quality indexes. At the microbial level, the alpha diversity was significantly higher in the MSTN -edited sheep ( P   〈  0.05), and the microbial composition was significantly different by PCoA analysis in the MSTN -edited and wild-type sheep. The abundance of Firmicutes significantly increased and Bacteroidota significantly decreased in the MSTN -edited sheep. At genus level, the abundance of Flavonifractor, Subdoligranulum, Ruthenibacterium, Agathobaculum, Anaerotignum, Oribacterium and Lactobacillus were significantly increased in the MSTN -edited sheep ( P   〈  0.05). Further analysis of functional differences was found that the carotenoid biosynthesis was significantly increased and the peroxisome, apoptosis, ferroptosis, N-glycan biosynthesis, thermogenesis, and adipocytokines pathways were decreased in the MSTN -edited sheep ( P   〈  0.05). Moreover, carbohydrate-active enzymes (CAZymes) results certified the abundance of the GH13_39, GH4, GH137, GH71 and PL17 were upregulated, and the GT41 and CBM20 were downregulated in the MSTN -edited sheep ( P   〈  0.05). Conclusions Our study suggested that MSTN inactivation remarkably influenced the composition and potential function of hindgut microbial communities of the sheep, and significantly promoted growth performance without affecting meat quality.
    Type of Medium: Online Resource
    ISSN: 1471-2180
    URL: Article
    DOI: 10.1186/s12866-022-02687-8
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2022
    detail.hit.zdb_id: 2041505-9
    SSG: 12
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  • 7
    Online Resource
    Online Resource
    Research on the collapse pressure of an elliptical wellbore considering the effect of weak planes
    Informa UK Limited ; 2020
    In:  Energy Sources, Part A: Recovery, Utilization, and Environmental Effects Vol. 42, No. 17 ( 2020-09-01), p. 2103-2119
    Details
    In: Energy Sources, Part A: Recovery, Utilization, and Environmental Effects, Informa UK Limited, Vol. 42, No. 17 ( 2020-09-01), p. 2103-2119
    Type of Medium: Online Resource
    ISSN: 1556-7036 , 1556-7230
    URL: Article
    DOI: 10.1080/15567036.2019.1607929
    Language: English
    Publisher: Informa UK Limited
    Publication Date: 2020
    detail.hit.zdb_id: 2238587-3
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  • 8
    Online Resource
    Online Resource
    Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system
    Springer Science and Business Media LLC ; 2015
    In:  Scientific Reports Vol. 5, No. 1 ( 2015-09-10)
    Details
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 5, No. 1 ( 2015-09-10)
    Abstract: Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of one-cell stage embryos with Cas9 mRNA and sgRNAs targeting two functional genes ( MSTN and FGF5 ), we successfully produced gene-modified goats with either one or both genes disrupted. The targeting efficiency of MSTN and FGF5 in cultured primary fibroblasts was as high as 60%, while the efficiency of disrupting MSTN and FGF5 in 98 tested animals was 15% and 21% respectively and 10% for double gene modifications. The on- and off-target mutations of the target genes in fibroblasts, as well as in somatic tissues and testis of founder and dead animals, were carefully analyzed. The results showed that simultaneous editing of several sites was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become a robust and efficient gene engineering tool in farm animals and therefore will be critically important and applicable for breeding.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    URL: Article
    DOI: 10.1038/srep13878
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2015
    detail.hit.zdb_id: 2615211-3
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  • 9
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    Online Resource
    Integrating miRNA and mRNA Expression Profiling Uncovers miRNAs Underlying Fat Deposition in Sheep
    Hindawi Limited ; 2017
    In:  BioMed Research International Vol. 2017 ( 2017), p. 1-11
    Details
    In: BioMed Research International, Hindawi Limited, Vol. 2017 ( 2017), p. 1-11
    Abstract: MicroRNAs (miRNAs) are endogenous, noncoding RNAs that regulate various biological processes including adipogenesis and fat metabolism. Here, we adopted a deep sequencing approach to determine the identity and abundance of miRNAs involved in fat deposition in adipose tissues from fat-tailed (Kazakhstan sheep, KS) and thin-tailed (Tibetan sheep, TS) sheep breeds. By comparing HiSeq data of these two breeds, 539 miRNAs were shared in both breeds, whereas 179 and 97 miRNAs were uniquely expressed in KS and TS, respectively. We also identified 35 miRNAs that are considered to be putative novel miRNAs. The integration of miRNA-mRNA analysis revealed that miRNA-associated targets were mainly involved in the gene ontology (GO) biological processes concerning cellular process and metabolic process, and miRNAs play critical roles in fat deposition through their ability to regulate fundamental pathways. These pathways included the MAPK signaling pathway, FoxO and Wnt signaling pathway, and focal adhesion. Taken together, our results define miRNA expression signatures that may contribute to fat deposition and lipid metabolism in sheep.
    Type of Medium: Online Resource
    ISSN: 2314-6133 , 2314-6141
    URL: Article
    DOI: 10.1155/2017/1857580
    Language: English
    Publisher: Hindawi Limited
    Publication Date: 2017
    detail.hit.zdb_id: 2698540-8
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  • 10
    Online Resource
    Online Resource
    A station location-allocation optimization problem in star-star pipeline network layout
    Informa UK Limited ; 2022
    In:  Petroleum Science and Technology
    Details
    In: Petroleum Science and Technology, Informa UK Limited
    Type of Medium: Online Resource
    ISSN: 1091-6466 , 1532-2459
    URL: Article
    DOI: 10.1080/10916466.2022.2147944
    Language: English
    Publisher: Informa UK Limited
    Publication Date: 2022
    detail.hit.zdb_id: 2490690-6
    detail.hit.zdb_id: 2043254-9
    SSG: 19,1
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