The generation and maintenance of genome edited zebrafish lines is typically labor intensive due to the lack of an easy visual read‐out for the modification. To facilitate this process, we have developed a novel method that relies on the inclusion of an artificial intron with a transgenic marker (InTraM) within the knock‐in sequence of interest, which upon splicing produces a transcript with a precise and seamless modification. We have demonstrated this technology by replacing the stop codon of the zebrafish fli1a gene with a transcriptional activator KALTA4, using an InTraM that enables red fluorescent protein expression in the heart.

中文翻译：

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带有转基因标记的内含子 (InTraM) 有助于内源基因报告系的高通量筛选。

基因组编辑斑马鱼线的生成和维护通常是劳动密集型的，因为缺乏简单的视觉读取来进行修改。为了促进这一过程，我们开发了一种新方法，该方法依赖于在感兴趣的敲入序列中包含带有转基因标记 (InTraM) 的人工内含子，该方法在剪接后产生具有精确和无缝修饰的转录本。我们已经通过使用转录激活因子 KALTA4替换斑马鱼fli1a基因的终止密码子证明了这项技术，使用 InTraM 可以在心脏中表达红色荧光蛋白。