Optogenetic in vivo cell manipulation in KillerRed-expressing zebrafish transgenics

BMC Dev Biol. 2010 Nov 2;10:110. doi: 10.1186/1471-213X-10-110.


Background: KillerRed (KR) is a novel photosensitizer that efficiently generates reactive oxygen species (ROS) in KR-expressing cells upon intense green or white light illumination in vitro, resulting in damage to their plasma membrane and cell death.

Results: We report an in vivo modification of this technique using a fluorescent microscope and membrane-tagged KR (mem-KR)-expressing transgenic zebrafish. We generated several stable zebrafish Tol2 transposon-mediated enhancer-trap (ET) transgenic lines expressing mem-KR (SqKR series), and mapped the transposon insertion sites. As mem-KR accumulates on the cell membrane and/or Golgi, it highlights cell bodies and extensions, and reveals details of cellular morphology. The photodynamic property of KR made it possible to damage cells expressing this protein in a dose-dependent manner. As a proof-of-principle, two zebrafish transgenic lines were used to affect cell viability and function: SqKR2 expresses mem-KR in the hindbrain rhombomeres 3 and 5, and elsewhere; SqKR15 expresses mem-KR in the heart and elsewhere. Photobleaching of KR by intense light in the heart of SqKR15 embryos at lower levels caused a reduction in pumping efficiency of the heart and pericardial edema and at higher levels - in cell death in the hindbrain of SqKR2 and in the heart of SqKR15 embryos.

Conclusions: An intense illumination of tissues expressing mem-KR affects cell viability and function in living zebrafish embryos. Hence, the zebrafish transgenics expressing mem-KR in a tissue-specific manner are useful tools for studying the biological effects of ROS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified* / anatomy & histology
  • Animals, Genetically Modified* / embryology
  • Animals, Genetically Modified* / genetics
  • Apoptosis
  • DNA Transposable Elements
  • Embryo, Nonmammalian* / anatomy & histology
  • Embryo, Nonmammalian* / drug effects
  • Embryo, Nonmammalian* / physiology
  • Enhancer Elements, Genetic
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / pharmacology*
  • Humans
  • In Situ Nick-End Labeling
  • Microscopy, Fluorescence / methods*
  • Morphogenesis / drug effects
  • Photosensitizing Agents / pharmacology*
  • Reactive Oxygen Species / pharmacology
  • Zebrafish* / anatomy & histology
  • Zebrafish* / embryology
  • Zebrafish* / genetics


  • DNA Transposable Elements
  • Photosensitizing Agents
  • Reactive Oxygen Species
  • killer red protein, Anthomedusae
  • Green Fluorescent Proteins