Compact engineered human mechanosensitive transactivation modules enable potent and versatile synthetic transcriptional control.
Compact engineered human mechanosensitive transactivation modules enable potent and versatile synthetic transcriptional control.
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DOI：
10.1038/s41592-023-02036-1
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发表时间：
2023-11
期刊：
影响因子：
48
通讯作者：
Hilton, Isaac B.
中科院分区：
文献类型：
--
作者：
Mahata, Barun;Cabrera, Alan;Brenner, Daniel A.;Guerra-Resendez, Rosa Selenia;Li, Jing;Goell, Jacob;Wang, Kaiyuan;Guo, Yannie;Escobar, Mario;Parthasarathy, Abinand Krishna;Szadowski, Hailey;Bedford, Guy;Reed, Daniel R.;Kim, Sunghwan;Hilton, Isaac B.
工程化的反式激活结构域（TADs）与可编程DNA结合平台相结合，彻底改变了合成转录控制。尽管最近在基于可编程CRISPR-Cas的反式激活（CRISPRa）技术方面取得了进展，但这些系统中使用的TAD通常含有耐受性差的元件和/或对于许多应用而言过大。在这里，我们定义和优化最小的TADs建立从人类机械敏感的转录因子。我们使用这些组件来构建有效且紧凑的多部分反式激活模块（MSN、NMS和eN 3x 9），并构建CRISPR-dCas 9募集增强激活模块（CRISPR-DREAM）平台。我们发现CRISPR-DREAM在哺乳动物细胞类型中具有特异性和稳健性，并有效地刺激了不同调控位点的转录。我们还表明MSN和NMS在I型、II型和V型CRISPR系统、转录激活因子样效应物和锌指蛋白中是可移植的。此外，作为概念的证明，我们使用dCas 9-NMS将人成纤维细胞有效地重编程为诱导的多能干细胞，并证明机械敏感性转录因子TADs在治疗上重要的原代人细胞类型中是有效的并且耐受良好。最后，我们利用这些工程化的TADs的紧凑和有效的特征来构建双重和一体化的CRISPRa AAV系统。总之，这些紧凑的人类TADs，融合模块和交付架构应该是有价值的生物医学应用中的合成转录控制。使用机械敏感性转录因子工程化的合成的基于CRISPR的反式激活结构域能够实现稳健的转录控制。
Engineered transactivation domains (TADs) combined with programmable DNA binding platforms have revolutionized synthetic transcriptional control. Despite recent progress in programmable CRISPR–Cas-based transactivation (CRISPRa) technologies, the TADs used in these systems often contain poorly tolerated elements and/or are prohibitively large for many applications. Here, we defined and optimized minimal TADs built from human mechanosensitive transcription factors. We used these components to construct potent and compact multipartite transactivation modules (MSN, NMS and eN3x9) and to build the CRISPR–dCas9 recruited enhanced activation module (CRISPR-DREAM) platform. We found that CRISPR-DREAM was specific and robust across mammalian cell types, and efficiently stimulated transcription from diverse regulatory loci. We also showed that MSN and NMS were portable across Type I, II and V CRISPR systems, transcription activator-like effectors and zinc finger proteins. Further, as proofs of concept, we used dCas9-NMS to efficiently reprogram human fibroblasts into induced pluripotent stem cells and demonstrated that mechanosensitive transcription factor TADs are efficacious and well tolerated in therapeutically important primary human cell types. Finally, we leveraged the compact and potent features of these engineered TADs to build dual and all-in-one CRISPRa AAV systems. Altogether, these compact human TADs, fusion modules and delivery architectures should be valuable for synthetic transcriptional control in biomedical applications. Synthetic CRISPR-based transactivation domains engineered using mechanosensitive transcription factors enable robust transcriptional control.
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影响因子：
64.8
作者：
Konermann S;Brigham MD;Trevino AE;Joung J;Abudayyeh OO;Barcena C;Hsu PD;Habib N;Gootenberg JS;Nishimasu H;Nureki O;Zhang F
通讯作者：
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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转录因子p300和MRTF-A协同增强MCF-7乳腺癌细胞迁移相关基因的表达
DOI：
10.1016/j.bbrc.2015.10.060
发表时间：
2015-11-27
影响因子：
3.1
作者：
He, Hongpeng;Wang, Dandan;Zhang, Tong-Cun
通讯作者：
Zhang, Tong-Cun
DOI：
10.1038/s41573-022-00476-6
发表时间：
2022-09
期刊：
Nature reviews. Drug discovery
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