当前位置: X-MOL 学术Cell Struct. Funct. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Biliverdin Reductase-A Deficiency Brighten and Sensitize Biliverdin-binding Chromoproteins.
Cell Structure and Function ( IF 2.0 ) Pub Date : 2020-08-21 , DOI: 10.1247/csf.20010
Kenju Kobachi 1 , Sota Kuno 2 , Shinya Sato 1 , Kenta Sumiyama 3 , Michiyuki Matsuda 1, 4 , Kenta Terai 1
Affiliation  

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.



中文翻译:



胆绿素还原酶 A 缺乏会使胆绿素结合色素蛋白变亮和敏感。



近红外 (NIR) 范围内的组织吸光度、光散射和自发荧光明显低于可见光范围。由于这些优点,近红外荧光蛋白 (FP)在体内成像方面的需求量很大。然而,由于 iRFP 等 NIR FP 在哺乳动物细胞中的暗淡,其应用仍然受到限制。与 GFP 及其变体相比,iRFP 需要胆绿素 (BV) 作为发色团。 iRFP 的暗淡至少部分是由于胆绿素还原酶 A (BLVRA) 快速降低 BV。在这里,我们建立了胆绿素还原酶 a敲除 ( Blvra –/– ) 小鼠,以增加细胞内 BV 浓度,从而增强 iRFP 荧光强度。正如预期的那样, Blvra –/–小鼠的所有检查组织中 iRFP 荧光强度均显着增加。同样,基于另一种 NIR FP mIFP 设计的基因编码钙指示剂 NIR-GECO1 在源自Blvra –/–小鼠的小鼠胚胎成纤维细胞中表现出荧光强度显着增加。我们将这种方法扩展到近红外光传感光遗传学工具 BphP1-PpsR2 系统,该系统也需要 BV 作为发色团。同样, Blvra基因的缺失显着增强了 HeLa 细胞的光反应。这些结果表明, Blvra –/–小鼠是近红外 FP 和近红外光传感光遗传学工具体内应用的多功能工具。

更新日期:2020-09-12
down
wechat
bug