Abstract

OTERO-ARROYO, RAFAEL; HERNANDEZ-HERRERA, DARWIN  and  PEREZ, JAIR. Production of bovine transgenic embryos by microinjection of a lentiviral vector pre and post fertilization. Rev.Bio.Agro [online]. 2020, vol.18, n.1, pp.64-73. ISSN 1692-3561.  https://doi.org/10.18684/bsaa.v18n1.1412.

The objective of this research was to compare the efficiency of microinjection of a lentiviral vector carrying the eGFP gene in mature oocytes (PRE) and zygotes (POS) six hours after in vitro fertilization (IVF). For this, 1777 mature oocytes were distributed in 7 treatments, one control (T1), three PRE (T2PRE, T3PRE and T4PRE) and three POS (T2POS, T3POS and T4POS) which were microinjected before (PRE) and after (POS) of IVF. The T4PRE and T4POS treatments were microinjected with the lentiviral vector. The blastocyst production rate at day eight and the expression of the eGFP protein in T4PRE and T4POS were evaluated. No differences were found in the blastocyst production rate between the T1 treatment (20,2%), the culture controls (T2PRE and T2POS) and the microinjection controls (T3PRE and T3POS). The presence of the lentiviral vector decreased (p<0,05) the blastocyst formation rate in the T4PRE and T4POS treatments (5,5 and 3,5%, respectively). All blastocysts produced in T4PRE and 71,4% produced in T4POS expressed the eGFP protein (p<0,05), mainly in the trophectoderm and in the internal cell mass of the blastocyst.

Keywords : Genetic modification; Green fluorescent protein; Transgenesis.

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