Lentiviral vectors are an effective method of introducing transgenes into the genome of early stage embryos because they transduce both dividing and non-dividing cells. Lentiviral pseudoparticles containing the coding sequence for the fluorescent protein DsRed were injected into freshly laid leopard gecko eggs. Tissue samples were collected from hatchlings, and the samples were tested for the presence of the transgene. Of the injected gecko population, greater than 89% of efficiency of transgenesis was confirmed using polymerase chain reaction (PCR). Histological evaluations revealed the presence of DsRed 2 in injected gecko organs; with protein production concentrated in the muscle, kidney, and heart. Therefore, lentiviral vectors appear to be viable technology to create transgenic geckos.
Restriction enzyme mediated integration (REMI) transgenesis and lentiviral transgenesis are effective methods of introducing transgenes into the genome of frogs. One aquatic amphibian species, Xenopus laevis, and one land dwelling species, Litoria caerulea, were chosen as subjects for transgenesis. REMI produced X. laevis that expressed the fluorescent protein DsRed. REMI was unsuccessful in producing transgenic Litoria. Therefore, lentiviral transgenesis was attempted. Hatchling Litoria tadpoles were exposed to replication defective lentiviral particles containing the coding sequence for DsRed. Histological evaluation revealed the presence of DsRed in brain, heart, liver, kidney, and muscle tissues. Therefore, lentiviral transgenesis appears to be a viable technique for producing transgenic land-dwelling frogs.
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