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Intron with transgenic marker ( InTraM ) facilitates high‐throughput screening of endogenous gene reporter lines
Author(s) -
Collins Russell T.,
Coxam Baptiste,
Fechner Ines,
Unterweger Iris,
Szymborska Anna,
Meier Katja,
Gerhardt Holger
Publication year - 2020
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.23391
Subject(s) - zebrafish , intron , rna splicing , biology , transgene , gene , green fluorescent protein , start codon , endogeny , genetics , microbiology and biotechnology , computational biology , messenger rna , rna , endocrinology
Summary 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.