Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ).
Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ).
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DOI：
10.1242/dmm.034561
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发表时间：
2018-09-27
影响因子：
4.3
通讯作者：
White RM
中科院分区：
文献类型：
--
作者：
Callahan SJ;Tepan S;Zhang YM;Lindsay H;Burger A;Campbell NR;Kim IS;Hollmann TJ;Studer L;Mosimann C;White RM
转基因动物对于癌症基因组学建模是非常宝贵的，但通常需要多个生殖系等位基因的复杂杂交以获得所需的组合。斑马鱼模型的优势在于转基因可以通过嵌合表达快速检测，但通常缺乏对肿瘤发作的空间和时间控制，这限制了它们在肿瘤进展和转移研究中的应用。为了克服这些限制，我们开发了一种称为成年斑马鱼转基因电穿孔（TEAZ）的方法。TEAZ可以递送具有感兴趣的启动子元件的DNA构建体，以在特定细胞类型中驱动荧光团、癌基因或基于CRISPR-Cas9的诱变盒。使用TEAZ，我们在空间受限的黑素细胞中通过BRAFV 600 E的背景下Cas9介导的Rb 1失活创建了高度侵袭性的黑色素瘤模型。与之前需要14个月才能开发的模型不同，我们发现TEAZ在17周内导致肿瘤发作，并且这些肿瘤在完全免疫活性的动物中发展。 由于所产生的肿瘤在高度确定的位置开始，我们可以通过荧光跟踪它们的进展，并记录到组织和转移性沉积物中的深度浸润。TEAZ可以使用合适的转基因启动子部署到其他组织和细胞类型，如心脏。TEAZ的多功能性使其可广泛用于体细胞基因改变和癌症进展的快速建模，其规模在其他体内系统中无法实现。总结：在这里，我们开发了成年斑马鱼转基因电穿孔（TEAZ），这是一种使研究人员能够通过将DNA元件局部引入成体组织来研究癌症和转移的方法。
Transgenic animals are invaluable for modeling cancer genomics, but often require complex crosses of multiple germline alleles to obtain the desired combinations. Zebrafish models have advantages in that transgenes can be rapidly tested by mosaic expression, but typically lack spatial and temporal control of tumor onset, which limits their utility for the study of tumor progression and metastasis. To overcome these limitations, we have developed a method referred to as Transgene Electroporation in Adult Zebrafish (TEAZ). TEAZ can deliver DNA constructs with promoter elements of interest to drive fluorophores, oncogenes or CRISPR-Cas9-based mutagenic cassettes in specific cell types. Using TEAZ, we created a highly aggressive melanoma model via Cas9-mediated inactivation of Rb1 in the context of BRAFV600E in spatially constrained melanocytes. Unlike prior models that take ∼4 months to develop, we found that TEAZ leads to tumor onset in ∼7 weeks, and these tumors develop in fully immunocompetent animals. As the resulting tumors initiated at highly defined locations, we could track their progression via fluorescence, and documented deep invasion into tissues and metastatic deposits. TEAZ can be deployed to other tissues and cell types, such as the heart, with the use of suitable transgenic promoters. The versatility of TEAZ makes it widely accessible for rapid modeling of somatic gene alterations and cancer progression at a scale not achievable in other in vivo systems. Summary: Here, we developed Transgene Electroporation in Adult Zebrafish (TEAZ), a method that enables researchers to study cancer and metastasis by introducing DNA elements focally into somatic adult tissue.
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影响因子：
7.3
作者：
Gao J;Aksoy BA;Dogrusoz U;Dresdner G;Gross B;Sumer SO;Sun Y;Jacobsen A;Sinha R;Larsson E;Cerami E;Sander C;Schultz N
通讯作者：
Schultz N
影响因子：
30.8
作者：
Krauthammer, Michael;Kong, Yong;Ha, Byung Hak;Evans, Perry;Bacchiocchi, Antonella;McCusker, James P.;Cheng, Elaine;Davis, Matthew J.;Goh, Gerald;Choi, Murim;Ariyan, Stephan;Narayan, Deepak;Dutton-Regester, Ken;Capatana, Ana;Holman, Edna C.;Bosenberg, Marcus;Sznol, Mario;Kluger, Harriet M.;Brash, Douglas E.;Stern, David F.;Materin, Miguel A.;Lo, Roger S.;Mane, Shrikant;Ma, Shuangge;Kidd, Kenneth K.;Hayward, Nicholas K.;Lifton, Richard P.;Schlessinger, Joseph;Boggon, Titus J.;Halaban, Ruth
通讯作者：
Halaban, Ruth
影响因子：
2.7
作者：
D'Agati G;Beltre R;Sessa A;Burger A;Zhou Y;Mosimann C;White RM
通讯作者：
White RM
DOI：
10.4049/jimmunol.1600959
发表时间：
2016-11-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Dee CT;Nagaraju RT;Athanasiadis EI;Gray C;Fernandez Del Ama L;Johnston SA;Secombes CJ;Cvejic A;Hurlstone AF
通讯作者：
Hurlstone AF
DOI：
10.1073/pnas.97.21.11403
发表时间：
2000-10-10
影响因子：
11.1
作者：
Kawakami, K;Shima, A;Kawakami, N
通讯作者：
Kawakami, N