DOI: 10.1073/pnas.2235688100. Electroporation and RNA interference in the rodent retina <i>in vivo</i> and <i>in vitro</i>

Electroporation and RNA interference in the rodent retina in vivo and in vitro

10.1073/pnas.2235688100

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The large number of candidate genes made available by comprehensive genome analysis requires that relatively rapid techniques for the study of function be developed. Here, we report a rapid and convenient electroporation method for both gain- and loss-of-function studies in vivo and in vitro in the rodent retina. Plasmid DNA directly injected into the subretinal space of neonatal rodent pups was taken up by a significant fraction of exposed cells after several pulses of high voltage. With this technique, GFP expression vectors were efficiently transfected into retinal cells with little damage to the operated pups. Transfected GFP allowed clear visualization of cell morphologies, and the expression persisted for at least 50 days. DNA-based RNA interference vectors directed against two transcription factors important in photoreceptor development led to photoreceptor phenotypes similar to those of the corresponding knockout mice. Reporter constructs carrying retinal cell type-specific promoters were readily introduced into the retina in vivo , where they exhibited the appropriate expression patterns. Plasmid DNA was also efficiently transfected into retinal explants in vitro by high-voltage pulses.

Bibliography

Matsuda, T., & Cepko, C. L. (2003). Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proceedings of the National Academy of Sciences, 101(1), 16â22.

Authors 2

Takahiko Matsuda (first)
Constance L. Cepko (additional)

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Dates

Type	When
Created	21 years, 7 months ago (Jan. 6, 2004, 1:12 p.m.)
Deposited	3 years, 4 months ago (April 25, 2022, 10:19 p.m.)
Indexed	3 days, 2 hours ago (Aug. 29, 2025, 6:15 a.m.)
Issued	21 years, 9 months ago (Nov. 5, 2003)
Published	21 years, 9 months ago (Nov. 5, 2003)
Published Online	21 years, 9 months ago (Nov. 5, 2003)
Published Print	21 years, 7 months ago (Jan. 6, 2004)

Funders 0

None

BibTeX

@article{Matsuda_2003, title={Electroporation and RNA interference in the rodent retina in vivo and in vitro}, volume={101}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.2235688100}, DOI={10.1073/pnas.2235688100}, number={1}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Matsuda, Takahiko and Cepko, Constance L.}, year={2003}, month=nov, pages={16–22} }

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