Combinatorial assembly of lumitoxins

David Nedrud, Daniel Schmidt

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Ion channels are among the most important proteins in neuroscience and serve as drug targets for many brain disorders. During development, learning, disease progression, and other processes, the activity levels of specific ion channels are tuned in a cell-type specific manner. However, it is difficult to assess how cell-specific changes in ion channel activity alter emergent brain functions. We have developed a protein architecture for fully genetically encoded light-activated modulation of endogenous ion channel activity. Fusing a genetically encoded photoswitch and an ion channel-modulating peptide toxin in a computationally designed fashion, this reagent, which we call Lumitoxins, can mediate light-modulation of specific endogenous ion channel activities in targeted cells. The modular lumitoxin architecture may be useful in a diversity of neuroscience tools. Here, we delineate how to construct lumitoxin genes from synthesized components, and provide a general outline for how to test their function in mammalian cell culture.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages193-209
Number of pages17
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1684
ISSN (Print)1064-3745

Keywords

  • Electrophysiology
  • Ion channel
  • LOV2
  • Optogenetic
  • Peptide toxin
  • Protein engineering

PubMed: MeSH publication types

  • Journal Article

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