Smith Sarah A, Lynch Kristen W
Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Methods Mol Biol. 2014;1126:243-55. doi: 10.1007/978-1-62703-980-2_18.
Cell-based splicing of minigenes is used extensively in the analysis of alternative splicing events. In particular, such assays are critical for identifying or confirming the in vivo relevance of cis- and trans-acting factors in the regulation of particular splicing patterns. Here we provide detailed information on the methods specific to the cell-based analysis of minigene splicing. In addition, we discuss some of the theoretical considerations that must be given to the design of the minigene and subsequent experimental conditions.
基于细胞的小基因剪接在可变剪接事件分析中被广泛应用。特别是,此类检测对于识别或确认顺式和反式作用因子在特定剪接模式调控中的体内相关性至关重要。在此,我们提供关于基于细胞的小基因剪接分析特定方法的详细信息。此外,我们还讨论了在小基因设计及后续实验条件设计时必须考虑的一些理论因素。