Journal
JOURNAL OF EUKARYOTIC MICROBIOLOGY
Volume 65, Issue 5, Pages 722-728Publisher
WILEY
DOI: 10.1111/jeu.12517
Keywords
Acidic compartmens; gene tagging; lysosomes; spaghetti monster tag; Trypanosoma cruzi
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Funding
- U.S. National Institutes of Health [AI107663]
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Membrane proteins in trypanosomatids are, in general, weakly expressed and confirmation of their subcellular localization frequently requires their overexpression with epitope tags. However, overexpression can lead to mislocalization of the probes. Viswanathan etal. (Nat. Methods, 2015, 12:568) described high performance tags for localization of weakly expressed proteins. We report here the use of these protein tags, named spaghetti monster, for CRISPR/Cas9-mediated C-terminal endogenous tagging of Trypanosoma cruzi to localize two weakly expressed transient receptor potential channels to acidic compartments. The results indicate that this method will improve the detection of membrane proteins in T.cruzi.
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