PUBLICATION
Intron with transgenic marker (InTraM) facilitates high-throughput screening of endogenous gene reporter lines
- Authors
- Collins, R.T., Coxam, B., Fechner, I., Unterweger, I., Szymborska, A., Meier, K., Gerhardt, H.
- ID
- ZDB-PUB-200813-7
- Date
- 2020
- Source
- Genesis (New York, N.Y. : 2000) 58(10-11): e23391 (Journal)
- Registered Authors
- Collins, Russell, Coxam, Baptiste, Fechner, Ines, Gerhardt, Holger, Meier, Katja, Szymborska-Mell, Anna
- Keywords
- CRISPR/Cas9, animal model, genome editing, screening, transgenesis, zebrafish
- MeSH Terms
-
- Gene Knock-In Techniques/methods*
- Animals
- CRISPR-Cas Systems
- Transgenes
- High-Throughput Screening Assays/methods*
- Transcription Factors/genetics
- Zebrafish
- Zebrafish Proteins/genetics
- Genes, Reporter*
- Gene Editing/methods*
- PubMed
- 32783355 Full text @ Genesis
Citation
Collins, R.T., Coxam, B., Fechner, I., Unterweger, I., Szymborska, A., Meier, K., Gerhardt, H. (2020) Intron with transgenic marker (InTraM) facilitates high-throughput screening of endogenous gene reporter lines. Genesis (New York, N.Y. : 2000). 58(10-11):e23391.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping