High-efficiency gene transfer into adult fish: a new tool to study fin regeneration.
Zebrafish represents an excellent model to study the function of vertebrate genes (e.g., well-developed genetics, large number of mutants, and genomic sequencing in progress), inasmuch as we have tools to manipulate gene expression. Recent use of injected morpholinos in eggs provides a good method to " knockdown " gene expression in early development (Nasevicius and Ekker, 2000), and the "caged" RNA injected in eggs allows to overexpress a gene in a specific set of cells (Ando et al., 2001). However, a method to specifically modify gene expression in the juvenile or in the adult is still missing. Such a method would be a very powerful tool to understand gene function in differentiated tissues. We describe here an electroporation-based approach, which allows gene transfer in adult tissues. Its efficiency was assessed using a GFP (green fluorescent protein) dependent assay. We then used this method to disrupt the Fgf signalling pathway during the process of regeneration.
Tawk
Marcel
M
Laboratoire de biologie moléculaire de la différenciation, Université Paris 7, Paris, France.
Tuil
David
D
Torrente
Yvan
Y
Vriz
Sophie
S
Paulin
Denise
D
D000818:Animals; D030801:Animals, Genetically Modified; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D012038:Regeneration / Q000502:physiology*; D015398:Signal Transduction; D015027:Zebrafish / Q000235:genetics* / Q000502:physiology
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins