Nutt et al (2001b)

Comparison of morpholino based translational inhibition during the development of Xenopus laevis and Xenopus tropicalis

S.L. Nutt, O. J. Bronchain, K. O. Hartley and E. Amaya

genesis. 30:110-113 (2001) (Full Article, 284K)

Abstract

Morpholino (MO) based inhibition of translational initiation represents an attractive methodology to eliminate gene

function during Xenopus development (Heasman, et al., 2000). However, the degree to which a given target protein

can be eliminated and the longevity of this effect during embryogenesis has not been documented. To examine the

efficacy of MOs, we have used transgenic Xenopus lines that harbour known numbers of integrations of a GFP

reporter under the control of the ubiquitous and highly expressed CMV promoter. In addition we have investigated the

longevity of the inhibitory effect by using transgenic lines expressing GFP specifically in the lens of tadpoles. These

transgenic lines represent the ideal control for the technique as the promoters arehighly expressed and GFP can be easily

detected by fluorescence and immunoblotting. Moreover, as GFP has no function in development, the levels of

inhibition can be tested in an otherwise normal individual. Here we report that MOs are able to efficiently and

specifically inhibit the translation of GFP in transgenic lines from Xenopus laevis and Xenopus tropicalis and the

inhibitory effect is long-lived, lasting into the tadpole stages.