5-Ph-IAA 是 IAA 的衍生物。 5-Ph-IAA 是一种配体，与 OsTIR1 (F74G) 突变体一起建立了生长素诱导型 degron 2 (AID2) 系统。 AID2 诱导 mAID 融合蛋白的快速有效消耗，以研究活细胞中的蛋白质功能，从而抑制肿瘤。
5-phenyl-indole-3-acetic acid (5-Ph-IAA) is a derivative of IAA. 5-Ph-IAA is a ligand to establish the auxin-induced degron 2 system (AID2) together with the OsTIR1 (F74G) mutant. Using AID2, degron fusion proteins can now be more precisely controlled, enabling target protein degradation with a half-life of 10 to 45 minutes by adding low doses of 5-Ph-IAA [1]. 5-Ph-IAA shows higher affinity to AtTIR1(F79G).
The mAID-fused targets were rapidly degraded by the addition of 1 μM 5-Ph-IAA and showed significant tumour suppression of mAID-BRD4 xenografts at all doses of 5-Ph-IAA treatment including 0, 1, 3, 10 mg/kg [1]. The treatment with 50 mM 5-Ph-IAA caused a rapid decrease in fluorescence intensity in many embryos (78%, 18/23 embryos), but 5 mM 5-Ph-IAA did not. 5-Ph-IAA can be used for loss-of-function experiments in C. elegans embryos, although treatment with 5-Ph-IAA is not always effective, possibly because the C. elegans embryo develops within the eggshell, which blocks its permeability to many compounds [2].
参考文献:
[1].Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat Commun. 2020 Nov 11;11(1):5701.
[2].Negishi T, Kitagawa S, Horii N, Tanaka Y, Haruta N, Sugimoto A, Sawa H, Hayashi KI, Harata M, Kanemaki MT. The auxin-inducible degron 2 (AID2) system enables controlled protein knockdown during embryogenesis and development in Caenorhabditis elegans. Genetics. 2022 Feb 4;220(2):iyab218.
5-phenyl-indole-3-acetic acid (5-Ph-IAA) 是 IAA 的衍生物。 5-Ph-IAA 是与 OsTIR1 (F74G) 突变体一起建立生长素诱导的 degron 2 系统 (AID2) 的配体。使用 AID2，现在可以更精确地控制 degron 融合蛋白，通过添加低剂量的 5-Ph-IAA [1] 使目标蛋白降解的半衰期为 10 至 45 分钟。 5-Ph-IAA对AtTIR1(F79G)具有更高的亲和力。
通过添加 1 μM 5-Ph-IAA，mAID 融合靶标会迅速降解，并且在所有剂量的 5-Ph-IAA 处理（包括 0、1、3、10毫克/千克 [1]。用 50 mM 5-Ph-IAA 处理导致许多胚胎（78%，18/23 胚胎）的荧光强度快速降低，但 5 mM 5-Ph-IAA 没有。 5-Ph-IAA 可用于秀丽隐杆线虫胚胎的功能丧失实验，尽管用 5-Ph-IAA 处理并不总是有效，这可能是因为秀丽隐杆线虫胚胎在蛋壳内发育，阻碍了其渗透性许多化合物 [2].
| Cell experiment [1]: | |
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Cell lines |
HCT116 cell |
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Preparation Method |
HCT116 cells were seeded at 1 × 105 cells/well in a six-well plate and grown for 2 days. Cells were treated with 0.5 μg/mL of doxycycline for 24 h, and then 1 μM 5-Ph-IAA was added. |
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Reaction Conditions |
1μM 5-Ph-IAA |
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Applications |
The mAID-fused targets were rapidly degraded by the addition of 1μM 5-Ph-IAA. |
| Animal experiment [2]: | |
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Animal models |
Balb/c-nu female(7 weeks old) weighing 16-20 g |
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Preparation Method |
Indicated HCT116 lines (1 × 10 5 cells for mAID-BRD4 and 2 × 10 5 cells for TOP2A-mAC) were resuspended in 0.1 ml of HBSS containing 0.05 ml of Matrigel. The suspension was injected into the both sides of flank. Six or 7 days after 5-Ph-IAA injection, the mice were randomized and treated daily with the indicated dose of 5-Ph-IAA by IP injection for additional 6 or 7 days. Tumour volume was measured on the indicated days. At the end of the experiment, xenograft tumour was removed and weighed. |
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Dosage form |
5-Ph-IAA by IP injection，0, 1, 3, 10 mg/kg |
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Applications |
significant tumour suppression of mAID-BRD4 xenografts at all doses of 5-Ph-IAA treatment |
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参考文献: [1]. Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat Commun. 2020 Nov 11;11(1):5701. |
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