Intron with transgenic marker (InTraM) facilitates high-throughput screening of endogenous gene reporter lines

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.