This is a preprint.
Comparative analysis of new, mScarlet-based red fluorescent tags in Caenorhabditis elegans
- PMID: 38915494
- PMCID: PMC11195195
- DOI: 10.1101/2024.06.11.598534
Comparative analysis of new, mScarlet-based red fluorescent tags in Caenorhabditis elegans
Update in
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Comparative analysis of new mScarlet-based red fluorescent tags in Caenorhabditis elegans.Genetics. 2024 Oct 7;228(2):iyae126. doi: 10.1093/genetics/iyae126. Genetics. 2024. PMID: 39103170 Free PMC article.
Abstract
One problem that has hampered the use of red fluorescent proteins in the fast-developing nematode C. elegans has been the substantial time delay in maturation of several generations of red fluorophores. The recently described mScarlet-I3 protein has properties that may overcome this limitation. We compare here the brightness and maturation time of CRISPR/Cas9 genome-engineered mScarlet, mScarlet3, mScarlet-I3 and GFP reporter knock-ins. Comparing the onset and brightness of expression of reporter alleles of C. elegans golg-4, encoding a broadly expressed Golgi resident protein, we found that the onset of detection of mScarlet-I3 in the embryo is several hours earlier than older versions of mScarlet and comparable to GFP. These findings were further supported by comparing mScarlet-I3 and GFP reporter alleles for pks-1, a gene expressed in the CAN neuron and cells of the alimentary system, as well as reporter alleles for the panneuronal, nuclear marker unc-75. Hence, the relative properties of mScarlet-I3 and GFP do not depend on cellular or subcellular context. In all cases, mScarlet-I3 reporters also show improved signal-to-noise ratio compared to GFP.
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References
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- Bindels D. S., Haarbosch L., van Weeren L., Postma M., Wiese K. E. et al., 2017. mScarlet: a bright monomeric red fluorescent protein for cellular imaging. Nature Methods 14: 53–56. - PubMed
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- Eroglu M., Yu B. and Derry W. B., 2023. Efficient CRISPR/Cas9 mediated large insertions using long single-stranded oligonucleotide donors in C. elegans. Febs j 290: 4429–4439. - PubMed
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