RRC ID 67757
Author Kobachi K, Kuno S, Sato S, Sumiyama K, Matsuda M, Terai K.
Title Biliverdin Reductase-A Deficiency Brighten and Sensitize Biliverdin-binding Chromoproteins.
Journal Cell Struct Funct
Abstract Tissue absorbance, light scattering, and autofluorescence are significantly lower in the near-infrared (NIR) range than in the visible range. Because of these advantages, NIR fluorescent proteins (FPs) are in high demand for in vivo imaging. Nevertheless, application of NIR FPs such as iRFP is still limited due to their dimness in mammalian cells. In contrast to GFP and its variants, iRFP requires biliverdin (BV) as a chromophore. The dimness of iRFP is at least partly due to rapid reduction of BV by biliverdin reductase-A (BLVRA). Here, we established biliverdin reductase-a knockout (Blvra-/-) mice to increase the intracellular BV concentration and, thereby, to enhance iRFP fluorescence intensity. As anticipated, iRFP fluorescence intensity was significantly increased in all examined tissues of Blvra-/- mice. Similarly, the genetically encoded calcium indicator NIR-GECO1, which is engineered based on another NIR FP, mIFP, exhibited a marked increase in fluorescence intensity in mouse embryonic fibroblasts derived from Blvra-/- mice. We expanded this approach to an NIR light-sensing optogenetic tool, the BphP1-PpsR2 system, which also requires BV as a chromophore. Again, deletion of the Blvra gene markedly enhanced the light response in HeLa cells. These results indicate that the Blvra-/- mouse is a versatile tool for the in vivo application of NIR FPs and NIR light-sensing optogenetic tools.Key words: in vivo imaging, near-infrared fluorescent protein, biliverdin, biliverdin reductase, optogenetic tool.
Volume 45(2)
Pages 131-141
Published 2020-8-21
DOI 10.1247/csf.20010
PMID 32581154
MeSH Animals Biliverdine / genetics Biliverdine / metabolism* Embryo, Mammalian / metabolism* Fibroblasts / metabolism* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* HeLa Cells Humans Mice Mice, Knockout Oxidoreductases Acting on CH-CH Group Donors / deficiency* Oxidoreductases Acting on CH-CH Group Donors / metabolism
IF 2.429
DNA material pCMV-VSV-G-RSV-Rev (RDB04393)