An nhr-23::mScarlet::3xMyc knock-in allele for studying spermatogenesis and molting.
An nhr-23::mScarlet::3xMyc knock-in allele for studying spermatogenesis and molting.
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DOI：
10.17912/micropub.biology.000996
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发表时间：
2023
影响因子：
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通讯作者：
Ward, Jordan D
中科院分区：
文献类型：
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作者：
Myles, Krista M;Ragle, James Matthew;Ward, Jordan D
线虫NHR-23是一种核激素受体转录因子，参与蜕皮、顶端细胞外基质结构和精子发生。为了确定NHR-23的表达动态，我们先前用GFP：：AID*：：3xFLAG标签标记了内源NHR-23基因座。为了使NHR-23与绿色荧光蛋白标记的目标因子共存，我们产生了一个携带mScarlet：：3xMyc标签的等效菌株，以产生C-末端融合。与GFP：：AID*：：3xFLAG类似，NHR-23：：mScarlet：：3xMyc在骨缝和皮下细胞、外阴前体细胞和生精胚系中表达。我们还观察到，当NHR-23不再定位于染色质时，NHR-23：：mScarlet在精子细胞和残留体中的表达呈弥漫性。对这一新定位的进一步研究可能有助于深入了解NHR-23对精子发生的调控。
C. elegans NHR-23 is a nuclear hormone receptor transcription factor involved in molting, apical extracellular matrix structure, and spermatogenesis. To determine NHR-23 expression dynamics, we previously tagged the endogenous nhr-23 locus with a GFP::AID*::3xFLAG tag. To allow co-localization of NHR-23 with green fluorescent protein-tagged factors of interest, we generated an equivalent strain carrying an mScarlet::3xMyc tag to produce a C-terminal fusion. Similar to the GFP::AID*::3xFLAG knock-in, NHR-23 ::mScarlet::3xMyc was expressed in seam and hypodermal cells, vulval precursor cells, and the spermatogenic germline. We also observed a diffuse NHR-23::mScarlet expression pattern in spermatids and residual bodies after NHR-23 ceased to localize on chromatin. Further examination of this novel localization may provide insight into NHR-23 regulation of spermatogenesis.