PUBLICATION

ΦC31 Integrase Mediates Efficient Cassette Exchange in the Zebrafish Germline

Authors
Hu, G., Goll, M.G., and Fisher, S.
ID
ZDB-PUB-110803-53
Date
2011
Source
Developmental dynamics : an official publication of the American Association of Anatomists   240(9): 2101-7 (Journal)
Registered Authors
Fisher, Shannon, Goll, Mary
Keywords
ΦC31 integrase, recombinase-mediated cassette exchange, zebrafish
MeSH Terms
  • Animals
  • Bacteriophages/enzymology*
  • Bacteriophages/genetics
  • DNA Nucleotidyltransferases/genetics
  • DNA Nucleotidyltransferases/metabolism*
  • Germ Cells
  • Integrases/genetics
  • Integrases/metabolism*
  • Models, Biological
  • Zebrafish
PubMed
21805532 Full text @ Dev. Dyn.
Abstract

Site-specific recombinases (SSRs) are powerful tools for genome manipulation, used in diverse organisms including Drosophila melanogaster, mouse, Arabidopsis, zebrafish, and human cultured cells. The integrase from the bacteriophage ΦC31 belongs to the large serine family of integrases, and in contrast to other widely used SSRs such as Cre and Flp, recombination is directional and therefore irreversible. We have developed a vector system for recombinase-mediated cassette exchange (RMCE) in the zebrafish, allowing swapping of the coding sequence in an integrated transgene. Utilizing codon-optimized ΦC31 integrase RNA bearing the 3′UTR from the nanos1 gene, we replaced the egfp coding sequence of an integrated reporter transgene with mCherry coding sequence. Recombination was achieved at high efficiency in both somatic cells and in the germline. We demonstrate an effective approach to RMCE, increasing the repertoire of tools available to manipulate the zebrafish genome.

Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping