In vivo single-cell electroporation for transfer of DNA and macromolecules

Jennifer E. Bestman, Rebecca C. Ewald, Shu Ling Chiu, Hollis T. Cline

Research output: Contribution to journalArticle

Abstract

Single-cell electroporation allows transfection of plasmid DNA or macrocmolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and combinations thereof. Experienced users can electroporate roughly 40 tadpoles per hour. The technique can be adapted for use with other charged transfer materials and in other systems and tissues where cells can be targeted with a micropipette. Under visual guidance, an electrode filled with transfer material is placed in a cell body-rich area of the tadpole brain and a train of voltage pulses applied, which electroporates a nearby cell. We show examples of successfully electroporated single cells, instances of common problems and troubleshooting suggestions. Single-cell electroporation is an affordable method to fluorescently label and genetically manipulate individual cells. This powerful technique enables observation of single cells in an otherwise normal environment.*Note: In the version of this article initially published online, the article's page numbers should have been 1267-1272. This error has been corrected in the PDF version of the article.

Original languageEnglish (US)
Pages (from-to)1267-1272
Number of pages6
JournalNature protocols
Volume1
Issue number3
DOIs
StatePublished - Aug 2006

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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