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

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Item Type:Article
Title:Intron with transgenic marker (InTraM) facilitates high-throughput screening of endogenous gene reporter lines
Creators Name:Collins, R.T. and Coxam, B. and Fechner, I. and Unterweger, I. and Szymborska, A. and Meier, K. and Gerhardt, H.
Abstract: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.
Keywords:Animal Model, CRISPR/Cas9, Genome Editing, Screening, Transgenesis, CRISPR-Cas Systems, Gene Editing, Gene Knock-In Techniques, Reporter Genes, High-Throughput Screening Assays, Transcription Factors, Transgenes, Animals, Zebrafish, Zebrafish Proteins / Genetics
Source:Genesis
ISSN:1526-954X
Publisher:Wiley
Volume:58
Number:10-11
Page Range:e23391
Date:November 2020
Official Publication:https://doi.org/10.1002/dvg.23391
PubMed:View item in PubMed

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