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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
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    Online Resource
    Transcriptome profile analysis of adipose tissues from fat and short-tailed sheep
    Elsevier BV ; 2014
    In:  Gene Vol. 549, No. 2 ( 2014-10), p. 252-257
    Details
    In: Gene, Elsevier BV, Vol. 549, No. 2 ( 2014-10), p. 252-257
    Type of Medium: Online Resource
    ISSN: 0378-1119
    URL: Article
    DOI: 10.1016/j.gene.2014.07.072
    RVK:
    WA 15000
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2014
    detail.hit.zdb_id: 1491012-3
    SSG: 12
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  • 9
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    Online Resource
    Comparative transcriptome analysis reveals potentially novel roles of Homeobox genes in adipose deposition in fat-tailed sheep
    Springer Science and Business Media LLC ; 2017
    In:  Scientific Reports Vol. 7, No. 1 ( 2017-11-03)
    Details
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 7, No. 1 ( 2017-11-03)
    Abstract: Adipose tissues are phenotypically, metabolically and functionally heterogeneous based on the sites of their deposition. Undesirable fat deposits in the body are often detrimental to animal and human health. To unravel the potential underlying mechanisms governing accumulation of adipose tissues in various regions of the body, i.e., subcutaneous (SAT), visceral (VAT) and tail (TAT), we profiled transcriptomes from Tan sheep, a Chinese indigenous breed with notable fat tail using RNA-seq. Upon comparison, we identified a total of 1,058 differentially expressed genes (DEGs) between the three adipose types (218, 324, and 795 in SAT/VAT, SAT/TAT, and VAT/TAT, respectively), from which several known key players were identified that are involved in lipid metabolic process, Wnt signals, Vitamin A metabolism, and transcriptional regulation of adipocyte differentiation. We also found that many elevated genes in VAT were notably enriched for key biological processes such as cytokine secretion, signaling molecule interaction and immune systems. Several developmental genes including HOXC11, HOXC12 and HOXC13, and adipose-expressed genes in the tail region, such as HOTAIR_2, HOTAIR_3 and SP9 were specially highlighted, indicating their strong associations with tail fat development in fat-tailed sheep. Our results provide new insight into exploring the specific fat deposition in tail, also contribute to the understanding of differences between adipose depots.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    URL: Article
    DOI: 10.1038/s41598-017-14967-9
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2017
    detail.hit.zdb_id: 2615211-3
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  • 10
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    Online Resource
    Whole-genome sequencing of eight goat populations for the detection of selection signatures underlying production and adaptive traits
    Springer Science and Business Media LLC ; 2016
    In:  Scientific Reports Vol. 6, No. 1 ( 2016-12-12)
    Details
    In: Scientific Reports, Springer Science and Business Media LLC, Vol. 6, No. 1 ( 2016-12-12)
    Abstract: The goat ( Capra hircus ) is one of the first farm animals that have undergone domestication and extensive natural and artificial selection by adapting to various environments, which in turn has resulted in its high level of phenotypic diversity. Here, we generated medium-coverage (9–13×) sequences from eight domesticated goat breeds, representing morphologically or geographically specific populations, to identify genomic regions representing selection signatures. We discovered ~10 million single nucleotide polymorphisms (SNPs) for each breed. By combining two approaches, ZH p and di values, we identified 22 genomic regions that may have contributed to the phenotypes in coat color patterns, body size, cashmere traits, as well as high altitude adaptation in goat populations. Candidate genes underlying strong selection signatures including coloration ( ASIP , KITLG , HTT , GNA11 , and OSTM1 ), body size ( TBX15 , DGCR8 , CDC25A , and RDH16 ), cashmere traits ( LHX2 , FGF9 , and WNT2 ), and hypoxia adaptation ( CDK2 , SOCS2 , NOXA1 , and ENPEP ) were identified. We also identified candidate functional SNPs within selected genes that may be important for each trait. Our results demonstrated the potential of using sequence data in identifying genomic regions that are responsible for agriculturally significant phenotypes in goats, which in turn can be used in the selection of goat breeds for environmental adaptation and domestication.
    Type of Medium: Online Resource
    ISSN: 2045-2322
    URL: Article
    DOI: 10.1038/srep38932
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2016
    detail.hit.zdb_id: 2615211-3
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