Rapid and efficient degradation of endogenous proteins in vivo identifies stage-specific roles of RNA Pol II pausing in mammalian development.
Rapid and efficient degradation of endogenous proteins in vivo identifies stage-specific roles of RNA Pol II pausing in mammalian development.
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DOI：
10.1016/j.devcel.2022.03.013
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发表时间：
2022-04-25
影响因子：
11.8
通讯作者：
Hadjantonakis, Anna-Katerina
中科院分区：
文献类型：
--
作者：
Abuhashem, Abderhman;Lee, Andrew S.;Joyner, Alexandra L.;Hadjantonakis, Anna-Katerina
靶向蛋白质降解方法为在各种模型系统中评估蛋白质的功能提供了一种独特的途径。最近，这些方法已经被应用于哺乳动物细胞培养模型，使得前所未有的对蛋白质功能的时间控制成为可能。然而，这些系统在体内组织和组织水平上的有效性还没有得到很好的证实。在这里，我们测试了降解标签(Dtag)降解系统在哺乳动物发育中的功能。我们产生了一只纯合子敲入小鼠，其FKBP12F36V标签与负延伸因子b(Nelfb)基因座融合，Nelfb基因是一种普遍表达的转录调节因子。在我们对哺乳动物发育和成年期的定向内源性蛋白质降解的验证中，我们证明了无论给药途径如何，Dtag系统在从受精卵到妊娠中期的胚胎中都是无毒、快速和有效的。此外，NELFB的急性耗尽在合子到2细胞的发育和合子基因组激活(ZGA)中显示了特定的作用。阿布哈希姆等人。在小鼠模型中评价Dtag degron系统的体内动力学和有效性。他们发现，该系统是安全的、可诱导的、高效的，并且在哺乳动物发育和成年组织中是可逆的。使用这个系统，他们确定了RNA Pol II暂停在调节哺乳动物受精卵基因组激活中的功能。
Targeted protein degradation methods offer a unique avenue to assess a protein’s function in a variety of model systems. Recently, these approaches have been applied to mammalian cell culture models, enabling unprecedented temporal control of protein function. However, the efficacy of these systems at the tissue and organismal levels in vivo is not well established. Here, we tested the functionality of the degradation tag (dTAG) degron system in mammalian development. We generated a homozygous knock-in mouse with a FKBP12F36V tag fused to Negative elongation factor b (Nelfb) locus, a ubiquitously expressed regulator of transcription. In our validation of targeted endogenous protein degradation across mammalian development and adulthood, we demonstrate that irrespective of the route of administration the dTAG system is non-toxic, rapid, and efficient in embryos from the zygote to mid-gestation stages. Additionally, acute depletion of NELFB revealed a specific role in zygote-to-2-cell development and Zygotic Genome Activation (ZGA). Abuhashem et al. evaluate the kinetics and efficacy of the dTAG degron system in vivo in a mouse model. They find that the system is safe, inducible, efficient, and reversable across mammalian development and adult tissues. Using this system, they identify a function for RNA Pol II pausing in regulating mammalian zygotic genome activation.
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影响因子：
1.5
作者：
Pimeisl, Inga-Marie;Tanriver, Yakup;Daza, Ray A.;Vauti, Franz;Hevner, Robert F.;Arnold, Hans-Henning;Arnold, Sebastian J.
通讯作者：
Arnold, Sebastian J.
DOI：
10.1007/978-1-61779-210-6_9
发表时间：
2011
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Piliszek, Anna;Kwon, Gloria S;Hadjantonakis, Anna-Katerina
通讯作者：
Hadjantonakis, Anna-Katerina
影响因子：
64.5
作者：
Rahl PB;Lin CY;Seila AC;Flynn RA;McCuine S;Burge CB;Sharp PA;Young RA
通讯作者：
Young RA
影响因子：
14.8
作者：
Nabet B;Roberts JM;Buckley DL;Paulk J;Dastjerdi S;Yang A;Leggett AL;Erb MA;Lawlor MA;Souza A;Scott TG;Vittori S;Perry JA;Qi J;Winter GE;Wong KK;Gray NS;Bradner JE
通讯作者：
Bradner JE
影响因子：
5.8
作者：
Abuhashem A;Garg V;Hadjantonakis AK
通讯作者：
Hadjantonakis AK