Methods of In Vivo Gene-Specific Knockdown Using Morpholino and Vivo-Morpholino Oligonucleotides

The functioning of the small GTPase gene, Ras-dva1 , has been studied during regeneration processes of the tadpole tails of the clawed frog Xenopus laevis . For this purpose, we have developed two approaches for the gene knockdown using injections of sequence-specific antisense morpholino oligonucle...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry 2018-05, Vol.44 (3), p.358-361
Main Authors: Ivanova, A. S., Korotkova, D. D., Martynova, N. Y., Averyanova, O. V., Zaraisky, A. G., Tereshina, M. B.
Format: Article
Language:English
Subjects:
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
Embryos
Gene expression
In vivo methods and tests
Letters to the Editor
Life Sciences
Oligonucleotides
Organic Chemistry
Regeneration
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Summary:The functioning of the small GTPase gene, Ras-dva1 , has been studied during regeneration processes of the tadpole tails of the clawed frog Xenopus laevis . For this purpose, we have developed two approaches for the gene knockdown using injections of sequence-specific antisense morpholino oligonucleotides (MO) or vivo-morpholino oligonucleotides (vivo-MO). It has been shown for the first time that intracellular Ras-dva -specific MO injected into Xenopus 4–16 of blastomere embryos or vivo-MO injected directly into the distal part of the tadpole tail at stages 40–41 efficiently inhibit the Ras-dva1 gene functioning and leads to the same morphological malformations of the tail regeneration. Using qRT-PCR, we detected the inhibition of expression of the regeneration marker genes ( Fgf20a and Msx1b ) on the first day after amputation in the tail tissues of tadpoles with the Ras-dva1 knockdown.
ISSN:1068-1620
1608-330X
DOI:10.1134/S106816201803007X