ArticlePloS one2012
ZyFISH: a simple, rapid and reliable zygosity assay for transgenic mice.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 7 citations in OpenAlex.
- Rapid and precise genotyping of transgene zygosity in mice using an allele-specific method.Life science alliance · 2023Article
- Mapping transgene insertion sites reveals the α-Cre transgene expression in both developing retina and olfactory neurons.Communications biology · 2022Article
- A neuroprotective role for microglia in prion diseases.The Journal of experimental medicine · 2016Article
- Simple and reliable genotyping protocol for mouse Prkdc(SCID) mutation.Journal of immunological methods · 2016Article
- Neurodegeneration and unfolded-protein response in mice expressing a membrane-tethered flexible tail of PrP.PloS one · 2015Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Microinjection of DNA constructs into fertilized mouse oocytes typically results in random transgene integration at a single genomic locus. The resulting transgenic founders can be used to establish hemizygous transgenic mouse lines. However, practical and experimental reasons often require that such lines be bred to homozygosity. Transgene zygosity can be determined by progeny testing assays which are expensive and time-consuming, by quantitative Southern blotting which is labor-intensive, or by quantitative PCR (qPCR) which requires transgene-specific design. Here, we describe a zygosity assessment procedure based on fluorescent in situ hybridization (zyFISH). The zyFISH protocol entails the detection of transgenic loci by FISH and the concomitant assignment of homozygosity using a concise and unbiased scoring system. The method requires small volumes of blood, is scalable to at least 40 determinations per assay, and produces results entirely consistent with the progeny testing assay. This combination of reliability, simplicity and cost-effectiveness makes zyFISH a method of choice for transgenic mouse zygosity determinations.
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Registered trials
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