RRC ID |
76491
|
Author |
Collins RT, Coxam B, Fechner I, Unterweger I, Szymborska A, Meier K, Gerhardt H.
|
Title |
Intron with transgenic marker (InTraM) facilitates high-throughput screening of endogenous gene reporter lines.
|
Journal |
Genesis
|
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.
|
Volume |
58(10-11)
|
Pages |
e23391
|
Published |
2020-11-1
|
DOI |
10.1002/dvg.23391
|
PMID |
32783355
|
MeSH |
Animals
CRISPR-Cas Systems
Gene Editing / methods*
Gene Knock-In Techniques / methods*
Genes, Reporter*
High-Throughput Screening Assays / methods*
Transcription Factors / genetics
Transgenes
Zebrafish
Zebrafish Proteins / genetics
|
IF |
1.76
|
Resource |
Zebrafish |
UAS:GFP |