Next-generation plasmids for transgenesis in zebrafish and beyond.
Next-generation plasmids for transgenesis in zebrafish and beyond.
复制标题
DOI：
10.1242/dev.201531
复制
发表时间：
2023-04-15
期刊：
影响因子：
4.6
通讯作者：
Mosimann, Christian
中科院分区：
文献类型：
--
作者：
Kemmler, Cassie L.;Moran, Hannah R.;Murray, Brooke F.;Scoresby, Aaron;Klem, John R.;Eckert, Rachel L.;Lepovsky, Elizabeth;Bertho, Sylvain;Nieuwenhuize, Susan;Burger, Sibylle;D'Agati, Gianluca;Betz, Charles;Puller, Ann-Christin;Felker, Anastasia;Ditrychova, Karolina;Botschi, Seraina;Affolter, Markus;Rohner, Nicolas;Ben Lovely, C.;Kwan, Kristen M.;Burger, Alexa;Mosimann, Christian
转基因是任何遗传模型的基本技术。基于Tol 2的转基因与Gateway兼容的载体集合配对，用可访问的模块化系统改变了斑马鱼转基因。在这里，我们建立了几个下一代转基因工具，斑马鱼和其他物种，以扩大和加强转基因应用。为了便于基因调控元件测试，我们产生了Gateway中间进入载体，其含有与几种荧光团CreERT 2和Gal 4偶联的小小鼠β-珠蛋白最小启动子。为了扩展转基因应用的色谱，我们建立了编码明亮的蓝色荧光蛋白mCerulean和mApple作为替代红色荧光团的中间入口载体。我们提出了一系列具有不同荧光团和亚细胞定位的基于p2 A肽的3′载体，以共标记表达感兴趣蛋白的细胞。最后，我们建立了Tol 2的目的载体携带斑马鱼exroh启动子驱动不同的荧光团作为松果体特异性的转基因标记，是活跃的孵化前，并通过成年。基于exorh的报告基因和转基因标记物也驱动无眼洞穴鱼（Astyanax）中的特定松果体表达。总之，我们的载体为斑马鱼、洞穴鱼和其他模型的转基因应用提供了多功能试剂。总结：几种新的质粒载体的产生，验证和应用，适用于斑马鱼和其他物种的转基因。
Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible modular system. Here, we establish several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse beta-globin minimal promoter coupled to several fluorophores, CreERT2 and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein mCerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3′ vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Finally, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker that is active before hatching and through adulthood. exorh-based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish (Astyanax). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish and other models. Summary: The generation, validation, and application of several new plasmid vectors suitable for transgenesis in zebrafish and other species.
登录
查看更多内容
影响因子：
2.7
作者：
Davison, Jon M.;Akitake, Courtney M.;Parsons, Michael J.
通讯作者：
Parsons, Michael J.
影响因子：
2.7
作者：
Akitake, Courtney M.;Macurak, Michelle;Goll, Mary G.
通讯作者：
Goll, Mary G.
DOI：
10.1073/pnas.2000942117
发表时间：
2020-12-01
影响因子：
11.1
作者：
Campbell BC;Nabel EM;Murdock MH;Lao-Peregrin C;Tsoulfas P;Blackmore MG;Lee FS;Liston C;Morishita H;Petsko GA
通讯作者：
Petsko GA
影响因子：
2.7
作者：
Davidson, AE;Balciunas, D;Ekker, SC
通讯作者：
Ekker, SC
影响因子：
2.7
作者：
AMSTERDAM, A;LIN, S;HOPKINS, N
通讯作者：
HOPKINS, N