Data/hora: 08/06/2024 20:59:06

Biblioteca(s): | Embrapa Gado de Leite.
Data corrente: | 14/01/2020
Data da última atualização: | 06/02/2024
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.
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.
Título: | Disulphide-less crotamine is effective for formation of DNA-peptide complex but is unable to improve bovine embryo transfection.
Ano de publicação: | 2020
Fonte/Imprenta: | Zygote, v. 28, n. 1, p. 72-79, 2020.
DOI: | https://doi.org/10.1017/s0967199419000716
Idioma: | Inglês
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.
Palavras-Chave: | Gene delivery; Microinjection; Nucleic acid-peptide complex.
Thesaurus Nal: | Embryotoxicity; Transgenesis.
Categoria do assunto: | --
Registro original: | Embrapa Gado de Leite (CNPGL)
Exemplares: | CNPGL - (24978-1) - UPC - AP - DD -  -  -  - DIS