Sperm-mediated gene transfer: effect on bovine in vitro embryo production

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dc.contributor.authorSimoes, Renata
dc.contributor.authorNicacio, Alessandra Corallo
dc.contributor.authorBinelli, Mario
dc.contributor.authorPaula-Lopes, Fabiola Freitas de
dc.contributor.authorMilazzotto, Marcella Pecora
dc.contributor.authorVisintin, Jose Antonio
dc.contributor.authorOrtiz D'Avila Assumpcao, Mayra Elena
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Federal do ABC (UFABC)
dc.contributor.institutionEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:34:41Z
dc.date.available2016-01-24T14:34:41Z
dc.date.issued2013-11-01
dc.description.abstractThe technique of sperm-mediated gene transfer (SMGT) can be used to delivery exogenous DNA into the oocyte. However, it has low repeatability and produces inconsistent results. in order to optimize this technique, it is necessary to study the mechanism by which DNA enters the sperm cell and integrates in the sperm genome. Furthermore, studies must focus in the maintenance of sperm cell viability and function. the aim of this study was to evaluate different SMGT protocols of sperm electroporation or capacitation (CaI) aiming to maintain sperm viability in the production of bovine embryos in vitro. Frozen-thawed semen was divided in two experimental groups (electroporation or CaI) and one control group (non-treated cells). for the electroporation method, five different voltages (100, 500, 750, 1000 or 1500 V) with 25 mu F capacitance were used. for CaI treatment, combinations of two CaI concentrations (250 nM or 500 nM), two incubation periods of sperm cells with CaI (1 or 5 min) and two incubation periods that mimicked time of sperm cell interaction with exogenous DNA molecules (1 or 2 h) were evaluated. According to our data, electroporation and CaI treatments do not prevent sperm penetration and oocyte fertilization and can be an alternative method to achieve satisfactory DNA delivery in SMGT protocols.en
dc.description.affiliationUniv São Paulo, Sch Vet Med & Anim Sci, Dept Anim Reprod, BR-05508270 São Paulo, Brazil
dc.description.affiliationFed Univ ABC UFABC, Ctr Ciencias Nat & Humanas, Santo Andre, SP, Brazil
dc.description.affiliationEmbrapa Beef Cattle Res Ctr, Campo Grande, MS, Brazil
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biol Sci, Diadema, SP, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biol Sci, Diadema, SP, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipIDFAPESP: 03/08542-5
dc.format.extent325-329
dc.identifierhttp://dx.doi.org/10.1017/S0967199412000147
dc.identifier.citationZygote. New York: Cambridge Univ Press, v. 21, n. 4, p. 325-329, 2013.
dc.identifier.doi10.1017/S0967199412000147
dc.identifier.issn0967-1994
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/36955
dc.identifier.wosWOS:000325654500002
dc.language.isoeng
dc.publisherCambridge Univ Press
dc.relation.ispartofZygote
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://journals.cambridge.org/action/displaySpecialPage?pageId=4676
dc.subjectBovineen
dc.subjectCalcium ionophoreen
dc.subjectElectroporationen
dc.subjectSMGTen
dc.subjectIVP embryoen
dc.titleSperm-mediated gene transfer: effect on bovine in vitro embryo productionen
dc.typeinfo:eu-repo/semantics/article
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