Comparative analysis of new mScarlet-based red fluorescent tags in Caenorhabditis elegans.

Autor:	Cao WX; Department of Biological Sciences, Columbia University, Howard Hughes Medical Institute, New York, NY 10027, USA., Merritt DM; Department of Biological Sciences, Columbia University, Howard Hughes Medical Institute, New York, NY 10027, USA., Pe K; Department of Biological Sciences, Columbia University, Howard Hughes Medical Institute, New York, NY 10027, USA., Cesar M; Department of Biological Sciences, Columbia University, Howard Hughes Medical Institute, New York, NY 10027, USA., Hobert O; Department of Biological Sciences, Columbia University, Howard Hughes Medical Institute, New York, NY 10027, USA.
Jazyk:	angličtina
Zdroj:	Genetics [Genetics] 2024 Oct 07; Vol. 228 (2).
DOI:	10.1093/genetics/iyae126
Abstrakt:	One problem that has hampered the use of red fluorescent proteins in the fast-developing nematode Caenorhabditis 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 onset of expression 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 pan-neuronal, 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. Competing Interests: Conflicts of interest The author(s) declare no conflicts of interest. (© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America.)
Databáze:	MEDLINE
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