Title | Monomerization of far-red fluorescent proteins. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | Wannier, Timothy M., Sarah K. Gillespie, Nicholas Hutchins, Scott R McIsaac, Sheng-Yi Wu, Yi Shen, Robert E. Campbell, Kevin S. Brown, and Stephen L. Mayo |
Journal | Proc Natl Acad Sci U S A |
Volume | 115 |
Issue | 48 |
Pagination | E11294-E11301 |
Date Published | 2018 11 27 |
ISSN | 1091-6490 |
Abstract | -class red fluorescent proteins (RFPs) are frequently used as biological markers, with far-red (λ ∼ 600-700 nm) emitting variants sought for whole-animal imaging because biological tissues are more permeable to light in this range. A barrier to the use of naturally occurring RFP variants as molecular markers is that all are tetrameric, which is not ideal for cell biological applications. Efforts to engineer monomeric RFPs have typically produced dimmer and blue-shifted variants because the chromophore is sensitive to small structural perturbations. In fact, despite much effort, only four native RFPs have been successfully monomerized, leaving the majority of RFP biodiversity untapped in biomarker development. Here we report the generation of monomeric variants of HcRed and mCardinal, both far-red dimers, and describe a comprehensive methodology for the monomerization of red-shifted oligomeric RFPs. Among the resultant variants is mKelly1 (emission maximum, λ = 656 nm), which, along with the recently reported mGarnet2 [Matela G, et al. (2017) 53:979-982], forms a class of bright, monomeric, far-red FPs. |
DOI | 10.1073/pnas.1807449115 |
Alternate Journal | Proc. Natl. Acad. Sci. U.S.A. |
PubMed ID | 30425172 |
PubMed Central ID | PMC6275547 |
Grant List | R21 EB018579 / EB / NIBIB NIH HHS / United States |