期刊
BIOCHEMICAL JOURNAL
卷 418, 期 -, 页码 567-574出版社
PORTLAND PRESS LTD
DOI: 10.1042/BJ20081949
关键词
far-red fluorescence; fluorescent protein; fusion protein; photostability; whole-body imaging
资金
- Howard Hughes Medical Institute [55005618, 55005630] Funding Source: Medline
- NIGMS NIH HHS [R01 GM070358, R01 GM073913, R01 GM070358-05, GM 070358, GM 073913, R01 GM073913-04] Funding Source: Medline
A vast colour palette of monomeric fluorescent proteins has been developed to investigate protein localization, motility and interactions. However, low brightness has remained a problem in far-red variants, which hampers multicolour labelling and whole-body imaging techniques. In the present paper, we report mKate2, a monomeric far-red fluorescent protein that is almost 3-fold brighter than the previously reported mKate and is 10-fold brighter than mPlum. The high-brightness, far-red emission spectrum, excellent pH resistance and photostability, coupled with low toxicity demonstrated in transgenic Xenopus laevis embryos, make mKate2 a superior fluorescent tag for imaging in living tissues. We also report tdKatushka2, a tandem far-red tag that performs well in fusions, provides 4-fold brighter near-IR fluorescence compared with mRaspberry or mCherry, and is 20-fold brighter than mPlum. Together, monomeric mKate2 and pseudomonomeric tdKatushka2 represent the next generation of extra-bright far-red fluorescent probes offering novel possibilities for fluorescent imaging of proteins in living cells and animals.
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