Restoration of the Majority of the Visual Spectrum by Using Modified Volvox Channelrhodopsin-1

We previously showed that blind rats whose vision was restored by gene transfer of Chlamydomonas channelrhodopsin-2 (ChR2) could only detect wavelengths less than 540 nm because of the action spectrum of the transgene product. Volvox-derived channelrhodopsin-1, VChR1, has a broader spectrum than ChR...

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Bibliographic Details
Published in:Molecular therapy 2014-08, Vol.22 (8), p.1434-1440
Main Authors: Tomita, Hiroshi, Sugano, Eriko, Murayama, Namie, Ozaki, Taku, Nishiyama, Fumiaki, Tabata, Kitako, Takahashi, Maki, Saito, Takehiko, Tamai, Makoto
Format: Article
Language:English
Subjects:
Animals
Bioengineering
Blindness - metabolism
Blindness - therapy
Chlamydomonas - genetics
Dependovirus - genetics
Gene therapy
Genetic Therapy - methods
Genetic Vectors - administration & dosage
Genetic Vectors - therapeutic use
HEK293 Cells
Humans
Light emitting diodes
Neurosciences
Original
Photoreceptors
Plasma
Polymerase chain reaction
Proteins
Rats
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Retina
Retina - physiopathology
Rhodopsin - genetics
Rhodopsin - metabolism
University graduates
Volvox - genetics
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Summary:We previously showed that blind rats whose vision was restored by gene transfer of Chlamydomonas channelrhodopsin-2 (ChR2) could only detect wavelengths less than 540 nm because of the action spectrum of the transgene product. Volvox-derived channelrhodopsin-1, VChR1, has a broader spectrum than ChR2. However, the VChR1 protein was mainly localized in the cytoplasm and showed weak ion channel properties when the VChR1 gene was transfected into HEK293 cells. We generated modified Volvox channelrhodopsin-1 (mVChR1), which is a chimera of Volvox channelrhodopsin-1 and Chlamydomonas channelrhodopsin-1 and demonstrated increased plasma membrane integration and dramatic improvement in its channel properties. Under whole-cell patch clamp, mVChR1-expressing cells showed a photo-induced current upon stimulation at 468–640 nm. The evoked currents in mVChR1-expressing cells were ~30 times larger than those in VChR1-expressing cells. Genetically, blind rats expressing mVChR1 via an adeno-associated virus vector regained their visual responses to light with wavelengths between 468 and 640 nm and their recovered visual responses were maintained for a year. Thus, mVChR1 is a candidate gene for gene therapy for restoring vision, and gene delivery of mVChR1 may provide blind patients access to the majority of the visible light spectrum.
ISSN:1525-0016
1525-0024
DOI:10.1038/mt.2014.81