Pseudotyped baculovirus is an effective gene expression tool for studying molecular function during axolotl limb regeneration

Axolotls can regenerate complex structures through recruitment and remodeling of cells within mature tissues. Accessing the underlying mechanisms at a molecular resolution is crucial to understand how injury triggers regeneration and how it proceeds. However, gene transformation in adult tissues can...

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Bibliographic Details
Published in:Developmental biology 2018-01, Vol.433 (2), p.262-275
Main Authors: Oliveira, Catarina R., Lemaitre, Regis, Murawala, Prayag, Tazaki, Akira, Drechsel, David N., Tanaka, Elly M.
Format: Article
Language:English
Subjects:
Ambystoma mexicanum - genetics
Ambystoma mexicanum - physiology
Amputation
Animals
Forelimb - physiology
Gene Expression Profiling
Gene Expression Regulation
Genes, Reporter
Genes, Synthetic
Genetic Vectors - genetics
Hedgehog Proteins - genetics
Hedgehog Proteins - physiology
Homeodomain Proteins - physiology
Humans
Membrane Glycoproteins - physiology
Mesoderm - cytology
Nucleopolyhedrovirus - genetics
Recombinant Proteins - metabolism
Regeneration - genetics
Regeneration - physiology
Transduction, Genetic
Transgenes
Viral Envelope Proteins - physiology
Wound Healing - genetics
Wound Healing - physiology
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Summary:Axolotls can regenerate complex structures through recruitment and remodeling of cells within mature tissues. Accessing the underlying mechanisms at a molecular resolution is crucial to understand how injury triggers regeneration and how it proceeds. However, gene transformation in adult tissues can be challenging. Here we characterize the use of pseudotyped baculovirus (BV) as an effective gene transfer method both for cells within mature limb tissue and within the blastema. These cells remain competent to participate in regeneration after transduction. We further characterize the effectiveness of BV for gene overexpression studies by overexpressing Shh in the blastema, which yields a high penetrance of classic polydactyly phenotypes. Overall, our work establishes BV as a powerful tool to access gene function in axolotl limb regeneration. •Pseudotyped baculovirus (BV) efficiently transfects axolotl intact limb tissue.•High efficient transfection is achieved in limb blastema.•PRRX1+ and PAX7+ cells are efficiently transfected and participate in regeneration.•BV elicits early and strong transgene expression.•BV is a complementary tool to current gene transfer methods.
ISSN:0012-1606
1095-564X
DOI:10.1016/j.ydbio.2017.10.008