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1.
Proc Natl Acad Sci U S A ; 116(42): 20930-20937, 2019 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-31575742

RESUMO

In macrolecithal species, cryopreservation of the oocyte and zygote is not possible due to the large size and quantity of lipid deposited within the egg. For birds, this signifies that cryopreserving and regenerating a species from frozen cellular material are currently technically unfeasible. Diploid primordial germ cells (PGCs) are a potential means to freeze down the entire genome and reconstitute an avian species from frozen material. Here, we examine the use of genetically engineered (GE) sterile female layer chicken as surrogate hosts for the transplantation of cryopreserved avian PGCs from rare heritage breeds of chicken. We first amplified PGC numbers in culture before cryopreservation and subsequent transplantation into host GE embryos. We found that all hatched offspring from the chimera GE hens were derived from the donor rare heritage breed broiler PGCs, and using cryopreserved semen, we were able to produce pure offspring. Measurement of the mutation rate of PGCs in culture revealed that 2.7 × 10-10 de novo single-nucleotide variants (SNVs) were generated per cell division, which is comparable with other stem cell lineages. We also found that endogenous avian leukosis virus (ALV) retroviral insertions were not mobilized during in vitro propagation. Taken together, these results show that mutation rates are no higher than normal stem cells, essential if we are to conserve avian breeds. Thus, GE sterile avian surrogate hosts provide a viable platform to conserve and regenerate avian species using cryopreserved PGCs.


Assuntos
Animais Geneticamente Modificados/genética , Cruzamento/métodos , Galinhas/genética , Células Germinativas/citologia , Infertilidade/veterinária , Animais , Animais Geneticamente Modificados/fisiologia , Galinhas/fisiologia , Criopreservação , Diploide , Transferência Embrionária , Feminino , Edição de Genes , Engenharia Genética , Masculino
2.
Transgenic Res ; 28(Suppl 2): 87-92, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31321689

RESUMO

The chicken is an exemplar of efficient intensive animal agriculture and provides two valuable food products, chicken meat and eggs. Only aquaculture is better, by efficiency, but poultry is still top, by mass of animal protein produced as food in the global context. However this efficiency and intensive production comes with a number of challenges. Though the genetics of selective breeding have led to dramatic improvements in yield, efficiency and product quality, traits that relate to disease and welfare outcomes have not been so tractable. These two issues are major impacts to the industry in terms of production and in terms of public perception. Both transgenic technology and genome editing have clear potential for impact in these two important areas. The reproductive biology of birds requires techniques very specific to birds to achieve heritable (germline) edited traits. These are quite involved and, even though they are now well-defined and reliable, there is room for improvement and advances can be expected in the future. Currently the key targets for this technology are modifying chicken genes involved in virus-receptor interactions and cellular response involved in infection. For the egg industry the technology is being applied to the issue of sex-selection for layer hens (and the removal of males), removal of allergens from egg white and the tailoring of eggs system to enhance the yield of influenza vaccine doses. Regulation and trading of the animals generated, and resulting food products, will significantly impact the value and future development of genome editing for poultry.


Assuntos
Hipersensibilidade a Ovo/genética , Edição de Genes/métodos , Engenharia Genética , Aves Domésticas/genética , Agricultura , Animais , Cruzamento , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Humanos , Aves Domésticas/crescimento & desenvolvimento , Seleção Artificial
3.
Mamm Genome ; 28(7-8): 315-323, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28612238

RESUMO

The application of gene editing (GE) technology to create precise changes to the genome of bird species will provide new and exciting opportunities for the biomedical, agricultural and biotechnology industries, as well as providing new approaches for producing research models. Recent advances in modifying both the somatic and germ cell lineages in chicken indicate that this species, and conceivably soon other avian species, has joined a growing number of model organisms in the gene editing revolution.


Assuntos
Aves/genética , Edição de Genes , Genoma , Animais , Sistemas CRISPR-Cas , Edição de Genes/métodos , Engenharia Genética , Células Germinativas/metabolismo , Modelos Animais , Especificidade de Órgãos
4.
J Sci Food Agric ; 93(14): 3574-80, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23744813

RESUMO

BACKGROUND: There is increasing evidence that consumption of plant bioactives such as polyphenols and glucosinolates reduces cardiovascular disease risk and improves endothelial function. In the Black Sea area, a number of plants are consumed alone and as ingredients in traditional foods, and dill, nettle, kale, Sideritis and persimmon were identified as bioactive-rich traditional food plants. The present study investigated the effects of plant extracts on cellular markers of endothelial function (eNOS activation and expression and ET-1 secretion). RESULTS: Treatment of human umbilical vein endothelial cells with persimmon extract significantly increased Akt and eNOS phosphorylation and nitric oxide metabolites and significantly decreased secretion of ET-1 to the media after 24 h compared with a vehicle control (all P < 0.01). None of the other plant extracts significantly altered any markers of endothelial function. CONCLUSION: These findings suggest that persimmon fruit contains bioactives that can improve endothelial function via activation of eNOS and reduction in ET-1 secretion, but that dill, kale, Sideritis and nettle do not.


Assuntos
Diospyros/química , Endotelina-1/metabolismo , Endotélio Vascular/efeitos dos fármacos , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico/metabolismo , Extratos Vegetais/farmacologia , Anethum graveolens/química , Mar Negro , Brassica/química , Bulgária , Cultura , Meios de Cultivo Condicionados/química , Endotélio Vascular/metabolismo , Ativação Enzimática/efeitos dos fármacos , Alimentos , Frutas , República da Geórgia , Células Endoteliais da Veia Umbilical Humana , Humanos , Nitratos/análise , Nitritos/análise , Fenóis/análise , Romênia , Federação Russa , Sérvia , Sideritis/química , Turquia , Ucrânia , Urtica dioica/química
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