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N-terminally endogenous mKate2-tagged HIM-4 in Caenorhabditis elegans.

PMID: 33937723 (view PubMed database entry)
DOI: 10.17912/micropub.biology.000386 (read at publisher's website )
PMCID: PMC8085682 (free full text version available)

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Shinji Ihara,

The gene <i>him-4</i> encodes the <i>C. elegans</i> homolog of hemicentin, an evolutionarily conserved extracellular matrix protein. Despite being an extracellular matrix protein, mutations in him-4 result in pleiotropic defects during development and demonstrate tissue fragility. While previous studies in <i>C. elegans</i> have confirmed the localization of either transgenic HIM-4::GFP or endogenous C-terminally tagged HIM-4::mNeonGreen, only green or green/yellow fluorescent protein-tagged HIM-4 are available to the C. elegans community. Here, I used CRISPR/Cas9 technology to insert the far-red fluorescence protein mKate2 at the <i>him-4</i> genomic locus, and established worms expressing mKate2::HIM-4. As expected, localization of mkate2::HIM-4 at the rachis was observed at the L4 stage. In contrast to the localization of type IV collagen or nidogen such as major components of the basement membrane, mkate2::HIM-4 was polarized at the anterior part of the pharyngeal basement membrane. A unique polarized localization pattern of pharyngeal basement membrane is maintained throughout the L1-L4 stages.

MicroPubl Biol (microPublication biology)
[2021, 2021:]

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