Neuron
Volume 4, Issue 2, February 1990, Pages 203-214
Journal home page for Neuron

Article
Lipofection of cDNAs in the embryonic vertebrate central nervous system

https://doi.org/10.1016/0896-6273(90)90095-WGet rights and content

Abstract

Neurons from the embryonic brain of Xenopus were transfected in vivo with a vector expressing luciferase cDNA using a simple lipofection procedure. Luciferase activity was monitored quantitatively, and the protein was immunolocalized in whole-mount embryonic brains. Luciferase-expressing neurons were often intensely labeled, displaying a Golgi-like filling of their dendrites, axons, and growth cones. Luciferase expression could be targeted to the retina by simply removing the skin epidermis covering the area and exposing the whole embryo to the DNA-lipofectin mixture. Luciferase activity in transfected embryos rose to peak values during the first 48 hr posttransfection and was still detectable 28 days later. Cotransfection experiments in which embryonic nervous tissue was exposed simultaneously to two different genes, luciferase and chloramphenicol acetyltransferase, showed that transfected cells coexpressed the two genes at an extremely high frequency (85%–100%). This offers the possibility of targeting functionally significant genes along with benign reporter genes in the developing CNS.

References (32)

  • J.R. de Wet et al.

    Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli

  • J.R. de Wet et al.

    Firefly luciferase gene: structure and expression in mammalian cells

    Mol. Cell. Biol.

    (1987)
  • L.D. Etkin et al.

    Distribution, expression and germ line transmission of exogenous DNA sequences following microinjection into Xenopus laevis eggs

    Development

    (1987)
  • R. Feiler et al.

    Targeted misexpression of a Drosophila opsin gene leads to altered visual function

    Nature

    (1988)
  • P.L. Felgner et al.

    Cationic liposome-mediated transfection

    Nature

    (1989)
  • P.L. Felgner et al.

    Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure

  • Cited by (0)

    View full text