title: Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong and Golic".
abstract: We have investigated the practicality of implementing a strategy for site-specific editing by homologous recombination in zebrafish analogous to that developed by Rong and Golic (Rong and Golic in Genetics 157:1307-1312, 2001) in Drosophila melanogaster. We analysed approximately 7,300 offspring from 22 crosses and demonstrated successful excision of the gene editing construct but failed to detect either gene editing or the random integration of the intact  editing construct subsequent to excision. The clustering of events in our data set demonstrates that the excision events are not occurring independently and emphasise that a promoter driving high level, tissue-specific transcription in meiotic cells is likely to be necessary if this general approach to site-specific editing by homologous recombination is to fulfil its potential.
authors.affiliation: School of Biology, Queens Medical Centre, Nottingham, NG2 5AN, UK.
authors.forename: Rosalind
authors.initials: R
authors.lastname: Brookfield
authors.forename: Felix
authors.initials: F
authors.lastname: Dafhnis-Calas
authors.forename: Zhengyao
authors.initials: Z
authors.lastname: Xu
authors.forename: William
authors.initials: W
authors.lastname: Brown
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D003433:Crosses, Genetic; D053903:DNA Breaks, Double-Stranded; D003851:Deoxyribonucleases / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D056426:Genetic Loci; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D059765:Homologous Recombination; D008540:Meiosis; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D019076:Transgenes; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D003851:Deoxyribonucleases