使用双重组酶马赛克分析（MADR）制备、组装和转导转基因元件。
Preparation, Assembly, and Transduction of Transgenic Elements Using Mosaic Analysis with Dual Recombinases (MADR).
机构信息
Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
出版信息
STAR Protoc. 2020 Dec 9;1(3):100199. doi: 10.1016/j.xpro.2020.100199. eCollection 2020 Dec 18.
This protocol focuses on the cloning and stable integration of sequences of interest by the use of a mosaic analysis with dual recombinases (MADR) plasmid that includes fusion proteins or independent proteins under the control of 2A peptide or IRES elements. Additionally, we describe how to generate a neural stem cell culture from Gt(ROSA)26SortJ mice, and validate the MADR plasmids and by neonatal mouse brain electroporation. This protocol can be generalized to analyze any transgenic element using MADR technology. For complete details on the use and execution of this protocol, please refer to Kim et al. (2019).
本方案重点介绍了利用包含融合蛋白或独立蛋白的嵌合分析双重组酶（MADR）质粒，通过 2A 肽或 IRES 元件的控制，对感兴趣的序列进行克隆和稳定整合。此外，我们还描述了如何从 Gt(ROSA)26SortJ 小鼠中生成神经干细胞培养物，并通过新生鼠脑电穿孔验证 MADR 质粒 和 。该方案可推广应用于使用 MADR 技术分析任何转基因元件。有关该方案使用和实施的完整详细信息，请参阅 Kim 等人。（2019 年）。
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