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dc.contributor.authorSOUZA, A. P. de
dc.contributor.authorBASTOS, A. P. A.
dc.contributor.authorFONSECA, F. M. da
dc.contributor.authorPANDOLFI, J. R. C.
dc.contributor.authorCOSTAMILAN, C. A. da V. L. R.
dc.contributor.authorMARQUES, M. G.
dc.date.accessioned2024-12-05T12:59:55Z-
dc.date.available2024-12-05T12:59:55Z-
dc.date.created2024-12-03
dc.date.issued2024
dc.identifier.citationAnimal Reproduction, v. 21, n. 4, e20240026, 2024.
dc.identifier.urihttp://www.alice.cnptia.embrapa.br/alice/handle/doc/1169957-
dc.descriptionAbstract: Polyethylenimine (PEI) has been explored as an efficient non-viral system for delivering genes to cells; however, there were no protocols for its use in porcine fetal fibroblasts (PFF). Therefore, we compared different concentrations of FITC-PEI (0.625, 1.25, 2.5, 5, 10, 20, 40, or 80 µg/mL) and incubation times (30 min, 1 h, or 2 h). It was observed that the incubation time did not affect the internalization of the PEI-FITC and that 30 min was sufficient to capture the complex. The concentrations higher than 10 µg/mL could reach many marked PFF (>90%). Then, two PEI concentrations were tested, 10 or 40 µg/mL, combined with an N/P of 2 with the pmhyGENIE-5 for 30 min. The percentage of PFF-GFP positive was similar between the PEI concentrations in the evaluation time points (24 h, 48 h, and 72 h). However, 40 µg/mL caused higher membrane damage rates. Thus, it can be concluded that concentrations between 10 – 80 µg/ml of PEI promote high incorporation rates, even in periods as short as 30 minutes. Furthermore, it can be stated that the transfection condition used in Polyplexes 1 (10 µg/mL of PEI and 37.5 µg/mL of pmhyGENIE5 for 30 min) efficiently produces genetically edited porcine fetal fibroblasts with low cell damage.
dc.language.isoeng
dc.rightsopenAccess
dc.subjectFibroblastos
dc.subjectTransfecção genética
dc.subjectPrimary fibroblast
dc.subjectCationic polymer
dc.subjectNon-viral vectors
dc.subjectGene delivery
dc.subjectNuclear transfer
dc.titlePolyethyleneimine-mediated gene transfection in porcine fetal fibroblasts.
dc.typeArtigo de periódico
dc.subject.thesagroGenética Animal
dc.subject.thesagroSuinocultura
dc.subject.thesagroFeto
dc.subject.nalthesaurusAnimal genetics
dc.subject.nalthesaurusSwine
riaa.ainfo.id1169957
riaa.ainfo.lastupdate2024-12-03
dc.identifier.doihttps://doi.org/10.1590/1984-3143-AR2024-0026
dc.contributor.institutionANDRESSA PEREIRA DE SOUZA, UNIVERSIDADE DO ESTADO DE SANTA CATARINA; ANA PAULA ALMEIDA BASTOS, CNPSA; FRANCISCO NOE DA FONSECA, GGINT; JOSE RODRIGO CLAUDIO PANDOLFI, CNPSA; CARLOS ANDRÉ DA VEIGA LIMA ROSA COSTAMILAN, UNIVERSIDADE DO ESTADO DE SANTA CATARINA; MARIANA GROKE MARQUES, CNPSA.
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