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Detail of Fig4C_N2_WT1

Kenju Kobachi, Sota Kuno, Shinya Sato, Kenta Sumiyama, Michiyuki Matsuda, Kenta Terai (2020) Biliverdin Reductase-A Deficiency Brighten and Sensitize Biliverdin-binding Chromoproteins., Cell structure and function, Volume 45, Number 2, pp. 131-141

Published in 2020 Aug 21 (Electronic publication in June 25, 2020, midnight )

• doi: 10.1247/csf.20010
• pmid: 32581154
  

(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.

• Animals
• Biliverdine/genetics
• Biliverdine/metabolism[major]
• Embryo, Mammalian/metabolism[major]
• Fibroblasts/metabolism[major]
• Green Fluorescent Proteins/genetics
• Green Fluorescent Proteins/metabolism[major]
• HeLa Cells
• Humans
• Mice
• Mice, Knockout
• Oxidoreductases Acting on CH-CH Group Donors/deficiency[major]
• Oxidoreductases Acting on CH-CH Group Donors/metabolism