Exogenous DNA length and quantity affect the transfection rate, but not sperm viability during sperm-mediated gene transfer

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dc.contributor(LBR) Lab. Biológicos Recombinantespt_BR
dc.contributor.authorFeitosa, Weber Beringuipt_BR
dc.contributor.authorMilazzotto, Marcella Pecorapt_BR
dc.contributor.authorMendes, Camilla Motapt_BR
dc.contributor.authorRocha, André Monteiro dapt_BR
dc.contributor.authorAvanzo, José Luispt_BR
dc.contributor.authorMartins, Elizabeth Angelica Lemept_BR
dc.contributor.authorVisintin, José Antoniopt_BR
dc.contributor.authorAssumpção, Mayra Elena Ortiz D'Ávilapt_BR
dc.date.accessioned2022-09-21T13:07:12Z-
dc.date.available2022-09-21T13:07:12Z-
dc.date.issued2022pt_BR
dc.identifier.citationFeitosa WB, Milazzotto MP, Mendes CM, Rocha AM, Avanzo JL, Martins EAL, et al. Exogenous DNA length and quantity affect the transfection rate, but not sperm viability during sperm-mediated gene transfer. Gene Rep. 2022 Dez; 29:101679. doi:10.1016/j.genrep.2022.101679.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/4515-
dc.description.abstractSpermatozoa have a spontaneous ability to take up exogenous DNA in a process regulated by specific mechanisms. This ability has been used to carry exogenous DNA into oocytes during fertilization to produce transgenic animals; a process called sperm-mediated gene transfer (SMGT). However, it is still an inefficient method and little is known about the effect of exogenous DNA once associated with spermatozoa, on sperm characteristics. Therefore, the objective of the present work was to evaluate the effects of exogenous DNA length and its amount on DNA uptake by bovine spermatozoa as well as spermatozoa viability. For that, spermatozoa (5 × 106 cells/mL) were incubated for 1 h at 38.5 °C with different exogenous DNA lengths (2.2, 5.5, or 8.5 kb) at different concentrations (number of molecules or ng). The association of exogenous DNA with spermatozoa was quantified by PCR real-time and the spermatozoa viability was evaluated by flow cytometry. Here, we show that no matter the amount of exogenous DNA used, larger sequences are less efficiently (p ˂ 0.05) associated with bovine spermatozoa. Besides that, the length and amount of exogenous DNA do not compromise sperm viability. Taken together, the results support that the length of exogenous DNA is more important than the amount used to influence its association with sperm cells. Thus, the size and quantity of exogenous DNA can be optimized to increase SMGT protocols, without altering the sperm viability.pt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.format.extent101679pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofGene Reportspt_BR
dc.rightsRestricted accesspt_BR
dc.titleExogenous DNA length and quantity affect the transfection rate, but not sperm viability during sperm-mediated gene transferpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1016/j.genrep.2022.101679pt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.identifier.citationvolume29pt_BR
dc.subject.keywordSMGTpt_BR
dc.subject.keywordgene transferpt_BR
dc.subject.keywordapoptosispt_BR
dc.subject.keywordsperm cellpt_BR
dc.subject.keywordtransgenicpt_BR
dc.relation.ispartofabbreviatedGene Reppt_BR
dc.identifier.citationabntv. 29, 101679, dez. 2022pt_BR
dc.identifier.citationvancouver2022 Dez; 29:101679pt_BR
dc.contributor.butantanMartins, Elizabeth Angelica Leme|:Pesquisador|:(LBR) Lab. Biológicos Recombinantespt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦03/10234-7pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦03/07456-8pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.grantfulltextembargo_20990101-
item.languageiso639-1English-
item.openairetypeArticle-
item.fulltextCom Texto completo-
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