A spontaneous TIR1 loss-of-function allele in C. elegans.
A spontaneous TIR1 loss-of-function allele in C. elegans.
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DOI：
10.17912/micropub.biology.000994
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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;Vo, An A;Ragle, James Matthew;Ward, Jordan D
生长素诱导降解决定子（AID）系统是一种广泛使用的条件性蛋白质去除系统。在实验过程中，我们耗尽了核激素受体转录因子NHR-23研究蜕皮，我们恢复了一个自发的抑制等位基因，绕过了L1幼虫逮捕所造成的NHR-23耗尽。这些突变体也未能耗尽菌株背景中的BFP：：AID报告基因，表明AID系统中存在更广泛的缺陷。这些动物携带在TIR 1中产生6个氨基酸重复的框内18个碱基对插入。在这些动物中的幼虫停滞可以通过表达来自染色体外阵列的野生型TIR 1转基因来恢复。失去这个阵列的姐妹兄弟姐妹在生长素上正常发育。总之，这些实验表明TIR 1突变导致nhr-23：：AID菌株的发育停滞丧失。这一结果强调了建立一个强大的二次筛选，以检测这样的突变体，如果执行正向遗传筛选与AID系统的重要性。
The auxin-inducible degron (AID) system is a widely-used system for conditional protein depletion. During the course of an experiment, we depleted the nuclear hormone receptor transcription factor NHR-23 to study molting, and we recovered a spontaneous suppressor allele that bypassed the L1 larval arrest caused by NHR-23 depletion. These mutants also failed to deplete a BFP::AID reporter in the strain background, suggesting a broader defect in the AID system. These animals carried an in-frame 18 base pair insertion that produced a 6 amino acid repeat in TIR1. The larval arrest in these animals could be restored by expressing a wild-type TIR1 transgene from an extrachromosomal array. Sister siblings that lost this array developed normally on auxin. Together, these experiments indicate that the TIR1 mutation was causing the loss of developmental arrest in the nhr-23::AID strain. This result highlights the importance of setting up a robust secondary screen to detect such mutants if performing forward genetic screens in conjunction with the AID system.