PUBLICATION
Homologous Recombination in Zebrafish ES Cells
- Authors
- Fan, L., Moon, J., Crodian, J., and Collodi, P.
- ID
- ZDB-PUB-060216-8
- Date
- 2006
- Source
- Transgenic Research 15(1): 21-30 (Journal)
- Registered Authors
- Collodi, Paul, Crodian, Jennifer, Fan, Lianchun, Moon, Jesung
- Keywords
- embryonic stem cells, homologous recombination, gene targeting, zebrafish
- MeSH Terms
-
- Myostatin/genetics
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- Animals
- Recombination, Genetic/genetics*
- Plasmids
- Zebrafish Proteins/genetics
- T-Box Domain Proteins/genetics
- Fetal Proteins
- Embryonic Stem Cells/metabolism*
- Cells, Cultured
- Gene Targeting
- Electroporation
- Animals, Genetically Modified
- Embryo, Nonmammalian/cytology
- PubMed
- 16475007 Full text @ Transgenic. Res.
Citation
Fan, L., Moon, J., Crodian, J., and Collodi, P. (2006) Homologous Recombination in Zebrafish ES Cells. Transgenic Research. 15(1):21-30.
Abstract
Targeted insertion of a plasmid by homologous recombination was demonstrated in zebrafish ES cell cultures. Two selection strategies were used to isolate ES cell colonies that contained targeted plasmid insertions in either the no tail or myostatin I gene. One selection strategy involved the manual isolation of targeted cell colonies that were identified by the loss of fluorescent protein gene expression. A second strategy used the diphtheria toxin A-chain gene in a positive-negative selection approach. Homologous recombination was confirmed by PCR, sequence and Southern blot analysis and colonies isolated using both selection methods were expanded and maintained for multiple passages. The results demonstrate that zebrafish ES cells have potential for use in a cell-mediated gene targeting approach.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping