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Highly Efficient Zebrafish Transgenesis Mediated by the Meganuclease I-SceI
- Source :
- Methods in Cell Biology, Methods in Cell Biology, Elsevier, 2004, 77, pp.381-401
- Publication Year :
- 2004
- Publisher :
- Elsevier, 2004.
-
Abstract
- Publisher Summary This chapter focuses on highly efficient zebrafish transgenesis mediated by the meganuclease I-SceI. Fish are excellent candidates for the production of transgenics for two important reasons. First, fish represent the largest and most diverse group of vertebrates and provide an advantageous system for in vivo studies of developmental processes to gain knowledge of gene regulation and the action of gene products in vertebrates. Second, conventional selective breeding of fish for improved growth or other characteristics is a very slow process. Transgenic fish technology has the potential to improve genetic traits such as increased growth potential, disease resistance, improved feed conversion efficiency, or other desirable genetic traits for aquaculture in one generation. The establishment of methods for successful transgenesis is one of the basic criteria for an organism to be referred to as model organism. Several endonucleases (meganucleases) encoded by introns and inteins have been shown to promote homing (lateral transfer) of their respective genetic elements into intron- or inteinless homologous allelic sites. By introducing sitespecific double-strand breaks (DSBs) in intronless alleles, these nucleases create recombinogenic ends that engage in gene conversion, resulting in duplication of the intron. The I-SceI system has been used as a tool in mammalian cells and Drosophila. The meganuclease can also be used in fish for comparative studies of cis-acting regulatory elements and homologous recombination (HR). Meganuclease transgenesis as described in this chapter could be further improved, including insulators upstream and downstream of the DNA of interest to protect the transgene from the influences of heterochromatin and epigenetic control of surrounding genomic sequences.
- Subjects :
- Microinjections
EENDODEOXYRIBONUCLEASE SCEI
[SDV]Life Sciences [q-bio]
Transgene
Animals Genetically Modified
ved/biology.organism_classification_rank.species
[INFO] Computer Science [cs]
MICROINJECTION
Deoxyribonucleases Type II Site-Specific
[INFO]Computer Science [cs]
[SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]
Gene conversion
GENE TRANSFER TECHNIQUE
Model organism
Zebrafish
ANIMAL GENETICALLY MODIFIED
Genetics
biology
ZEBRAFISH
ved/biology
Gene Transfer Techniques
Meganuclease I-SceI
biology.organism_classification
[SDV] Life Sciences [q-bio]
Transgenesis
Meganuclease
Homologous recombination
Subjects
Details
- ISSN :
- 0091679X
- Database :
- OpenAIRE
- Journal :
- Methods in Cell Biology, Methods in Cell Biology, Elsevier, 2004, 77, pp.381-401
- Accession number :
- edsair.doi.dedup.....12209f91a1d25a18c47ec992f61093b8