用于实时多色生物成像的明亮发光蛋白的五种颜色变体。,Nature Communicati
用于实时多色生物成像的明亮发光蛋白的五种颜色变体。
Nature Communications
(
IF
14.7
)
Pub Date : 2016-12-14
, DOI:
10.1038/ncomms13718
Kazushi Suzuki
1
,
Taichi Kimura
2
,
Hajime Shinoda
1
,
Guirong Bai
3
,
Matthew J Daniels
4
,
Yoshiyuki Arai
1,
2,
3
,
Masahiro Nakano
1,
2,
3
,
Takeharu Nagai
1,
2,
3
Affiliation
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
Department of Biotechnology, School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.
Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
BHF Centre for Regenerative Medicine, Division of Cardiovascular Medicine, West Wing Level 6, John Radcliffe Hospital, Oxford OX3 9DU, UK.
在不能应用荧光成像的情况下，陪护老人13825404095发光成像作为一种有前途的生物成像方式而受到关注。然而，由于缺乏在可见光谱范围内发射的明亮发光蛋白，限制了多色和动态成像的更广泛应用。在这里，我们报告了明亮发光蛋白的五种新光谱变体，增强型纳米灯 (eNL)，由最亮的荧光素酶 NanoLuc 与各种颜色的荧光蛋白连接而成。eNL 允许五色活细胞成像，以及检测单个蛋白质复合物甚至单个分子。我们还开发了一种基于 eNL 的 Ca 2+指示剂，具有 500% 的信号变化，可以对自发的 Ca 2+进行成像心肌细胞和神经细胞模型的动力学。这些 eNL 探针不仅有助于在活细胞中进行多色成像，而且还有助于对多种蛋白质进行灵敏成像，即使在非常低的表达水平下也是如此。
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Five colour variants of bright luminescent protein for real-time multicolour bioimaging.
Luminescence imaging has gained attention as a promising bio-imaging modality in situations where fluorescence imaging cannot be applied. However, wider application to multicolour and dynamic imaging is limited by the lack of bright luminescent proteins with emissions across the visible spectrum. Here we report five new spectral variants of the bright luminescent protein, enhanced Nano-lantern (eNL), made by concatenation of the brightest luciferase, NanoLuc, with various colour hues of fluorescent proteins. eNLs allow five-colour live-cell imaging, as well as detection of single protein complexes and even single molecules. We also develop an eNL-based Ca2+ indicator with a 500% signal change, which can image spontaneous Ca2+ dynamics in cardiomyocyte and neural cell models. These eNL probes facilitate not only multicolour imaging in living cells but also sensitive imaging of a wide repertoire of proteins, even at very low expression levels.