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 | Acesso ao texto completo restrito à biblioteca da Embrapa Gado de Leite. Para informações adicionais entre em contato com cnpgl.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Gado de Leite. |
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Data corrente: |
14/01/2020 |
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Data da última atualização: |
06/02/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
FREITAS, V. J. F.; CAMPELO, I. S.; SILVA, M. M. A. S.; CAVALCANTI, C. M.; TEIXEIRA, D. I. A.; CAMARGO, L. S. de A.; MELO L. M.; RÁDIS-BAPTISTA, G. |
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Afiliação: |
Vicente J. F. Freitas; Iana S. Campelo; Mirelly M .A. S. Silva; Camila M. Cavalcanti; Dárcio I. A. Teixeira; LUIZ SERGIO DE ALMEIDA CAMARGO, CNPGL; Luciana M. Melo; Gandhi Rádis-Baptista. |
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Título: |
Disulphide-less crotamine is effective for formation of DNA-peptide complex but is unable to improve bovine embryo transfection. |
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Ano de publicação: |
2020 |
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Fonte/Imprenta: |
Zygote, v. 28, n. 1, p. 72-79, 2020. |
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DOI: |
https://doi.org/10.1017/s0967199419000716 |
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Idioma: |
Inglês |
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Conteúdo: |
This study aimed to investigate the ability of disulphide-less crotamine (dLCr) to complex DNA and to evaluate whether the DNA-dLCr complex is capable of improving transfection in bovine embryos. Three experiments were performed to: (i) evaluate the formation and stability of the DNA-dLCr complex; (ii) assess the dLCr embryotoxicity by exposure of bovine embryos to dLCr; and (iii) assess the efficiency of bovine embryo transfection after microinjection of the DNA-dLCr complex or green fluorescent protein (GFP) plasmid alone (control). DNA complexation by dLCr after 30 min of incubation at 1:100 and 1:50 proportions presented higher efficiency (P < 0.05) than the two controls: native crotamine (NCr) 1:10 and lipofectamine. There was no difference between DNA-dLCr 1:25 and the controls. The DNA-dLCr complexation was evaluated at different proportions and times. In all, at least half of maximum complexation was achieved within the initial 30 min. No embryotoxicity of dLCr was verified after exposure of in vitro fertilized embryos to different concentrations of the peptide. The effectiveness of dLCr to improve exogenous gene expression was evaluated by microinjection of the DNA-dLCr complex into in vitro fertilized zygotes, followed by verification of both embryo development and GFP expression. From embryos microinjected with DNA only, 4.6% and 2.8% expressed the GFP transgene at day 5 and day 7, respectively. The DNA-dLCr complex did not increase the number of GFP-positive embryos. In conclusion, dLCr forms a complex with DNA and its application in in vitro culture is possible. However, the dLCr peptide sequence should be redesigned to improve GFP expression. MenosThis study aimed to investigate the ability of disulphide-less crotamine (dLCr) to complex DNA and to evaluate whether the DNA-dLCr complex is capable of improving transfection in bovine embryos. Three experiments were performed to: (i) evaluate the formation and stability of the DNA-dLCr complex; (ii) assess the dLCr embryotoxicity by exposure of bovine embryos to dLCr; and (iii) assess the efficiency of bovine embryo transfection after microinjection of the DNA-dLCr complex or green fluorescent protein (GFP) plasmid alone (control). DNA complexation by dLCr after 30 min of incubation at 1:100 and 1:50 proportions presented higher efficiency (P < 0.05) than the two controls: native crotamine (NCr) 1:10 and lipofectamine. There was no difference between DNA-dLCr 1:25 and the controls. The DNA-dLCr complexation was evaluated at different proportions and times. In all, at least half of maximum complexation was achieved within the initial 30 min. No embryotoxicity of dLCr was verified after exposure of in vitro fertilized embryos to different concentrations of the peptide. The effectiveness of dLCr to improve exogenous gene expression was evaluated by microinjection of the DNA-dLCr complex into in vitro fertilized zygotes, followed by verification of both embryo development and GFP expression. From embryos microinjected with DNA only, 4.6% and 2.8% expressed the GFP transgene at day 5 and day 7, respectively. The DNA-dLCr complex did not increase the number of GFP-positive embr... Mostrar Tudo |
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Palavras-Chave: |
Gene delivery; Microinjection; Nucleic acid-peptide complex. |
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Thesaurus Nal: |
Embryotoxicity; Transgenesis. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02565naa a2200277 a 4500
001 2118715
005 2024-02-06
008 2020 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1017/s0967199419000716$2DOI
100 1 $aFREITAS, V. J. F.
245 $aDisulphide-less crotamine is effective for formation of DNA-peptide complex but is unable to improve bovine embryo transfection.$h[electronic resource]
260 $c2020
520 $aThis study aimed to investigate the ability of disulphide-less crotamine (dLCr) to complex DNA and to evaluate whether the DNA-dLCr complex is capable of improving transfection in bovine embryos. Three experiments were performed to: (i) evaluate the formation and stability of the DNA-dLCr complex; (ii) assess the dLCr embryotoxicity by exposure of bovine embryos to dLCr; and (iii) assess the efficiency of bovine embryo transfection after microinjection of the DNA-dLCr complex or green fluorescent protein (GFP) plasmid alone (control). DNA complexation by dLCr after 30 min of incubation at 1:100 and 1:50 proportions presented higher efficiency (P < 0.05) than the two controls: native crotamine (NCr) 1:10 and lipofectamine. There was no difference between DNA-dLCr 1:25 and the controls. The DNA-dLCr complexation was evaluated at different proportions and times. In all, at least half of maximum complexation was achieved within the initial 30 min. No embryotoxicity of dLCr was verified after exposure of in vitro fertilized embryos to different concentrations of the peptide. The effectiveness of dLCr to improve exogenous gene expression was evaluated by microinjection of the DNA-dLCr complex into in vitro fertilized zygotes, followed by verification of both embryo development and GFP expression. From embryos microinjected with DNA only, 4.6% and 2.8% expressed the GFP transgene at day 5 and day 7, respectively. The DNA-dLCr complex did not increase the number of GFP-positive embryos. In conclusion, dLCr forms a complex with DNA and its application in in vitro culture is possible. However, the dLCr peptide sequence should be redesigned to improve GFP expression.
650 $aEmbryotoxicity
650 $aTransgenesis
653 $aGene delivery
653 $aMicroinjection
653 $aNucleic acid-peptide complex
700 1 $aCAMPELO, I. S.
700 1 $aSILVA, M. M. A. S.
700 1 $aCAVALCANTI, C. M.
700 1 $aTEIXEIRA, D. I. A.
700 1 $aCAMARGO, L. S. de A.
700 1 $aMELO L. M.
700 1 $aRÁDIS-BAPTISTA, G.
773 $tZygote$gv. 28, n. 1, p. 72-79, 2020.
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Embrapa Gado de Leite (CNPGL) |
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| 1. |  | PRATES, A.; QUADROS, L. F. P.; MACHADO, D.; BARRO, R.; CEZIMBRA, I. M.; BONNET, O. J. F.; GENRO, T. C. M.; CARVALHO, P. C. F. Grazing intensity as a strategy to mitigate methane in native grassland. In: INTERNATIONAL SYMPOSIUM ON GREENHOUSE GASES IN AGRICULTURE, 2., 2016, Campo Grande, MS. Proceedings... Brasília, DF: Embrapa, 2016. p. 205-212. (Embrapa Gado de Corte. Documentos, 216). Coordenador: Roberto Giolo de Almeida. II SIGEE.| Tipo: Artigo em Anais de Congresso |
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