PUBLICATION
Creation of zebrafish knock-in reporter lines in the nefma gene by Cas9-mediated homologous recombination
- Authors
- Eschstruth, A., Schneider-Maunoury, S., Giudicelli, F.
- ID
- ZDB-PUB-191002-4
- Date
- 2019
- Source
- Genesis (New York, N.Y. : 2000) 58(1): e23340 (Journal)
- Registered Authors
- Schneider-Maunoury, Sylvie
- Keywords
- CRISPR/Cas9, genome editing, knock-in, nefma, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- CRISPR-Cas Systems/genetics
- Gene Editing/methods
- Gene Knock-In Techniques/methods*
- Gene Targeting/methods
- Genetic Engineering/methods*
- Genome/genetics
- Homologous Recombination/genetics
- Intermediate Filaments/genetics
- RNA, Guide, Kinetoplastida/genetics
- Zebrafish/genetics
- PubMed
- 31571409 Full text @ Genesis
Citation
Eschstruth, A., Schneider-Maunoury, S., Giudicelli, F. (2019) Creation of zebrafish knock-in reporter lines in the nefma gene by Cas9-mediated homologous recombination. Genesis (New York, N.Y. : 2000). 58(1):e23340.
Abstract
CRISPR/Cas9-based strategies are widely used for genome editing in many organisms, including zebrafish. Although most applications consist in introducing double strand break (DSB)-induced mutations, it is also possible to use CRISPR/Cas9 to enhance homology directed repair (HDR) at a chosen genomic location to create knock-ins with optimally controlled precision. Here, we describe the use of CRISPR/Cas9-targeted DSB followed by HDR to generate zebrafish transgenic lines where exogenous coding sequences are added in the nefma gene, in frame with the endogenous coding sequence. The resulting knock-in embryos express the added gene (fluorescent reporter or KalTA4 transactivator) specifically in the populations of neurons that express nefma, making them convenient tools for research on these populations.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping