ArticlePloS one2013
A site-specific recombinase-based method to produce antibiotic selectable marker free transgenic cattle.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 32 citations in OpenAlex.
- Secretion of IFN-γ by Transgenic Mammary Epithelial CellsFrontiers in veterinary science · 2022Article
- Characterization of the regulatory 5'-flanking region of bovine mucin 2 (MUC2) gene.Molecular and cellular biochemistry · 2021Article
- Highly efficient CRISPR-targeting of the murine Hipp11 intergenic region supports inducible human transgene expression.Molecular biology reports · 2020Article
- Site-Specific Fat-1 Knock-In Enables Significant Decrease of n-6PUFAs/n-3PUFAs Ratio in Pigs.G3 (Bethesda, Md.) · 2018Article
- PhiC31-based Site-Specific Transgenesis System for Production of Transgenic Bovine Embryos by Somatic Cell Nuclear Transfer and Intracytoplasmic Sperm Injection.Cell journal · 2018Article
- ФC31 Integrase-Mediated Isolation and Characterization of Novel Safe Harbors for Transgene Expression in the Pig Genome.International journal of molecular sciences · 2018Article
- Cell-penetrating peptide-driven Cre recombination in porcine primary cells and generation of marker-free pigs.PloS one · 2018Article
- Blastocyst Formation Rate and Transgene Expression are Associated with Gene Insertion into Safe and Non-Safe Harbors in the Cattle Genome.Scientific reports · 2017Article
- Isozygous and selectable marker-free MSTN knockout cloned pigs generated by the combined use of CRISPR/Cas9 and Cre/LoxP.Scientific reports · 2016Article
- Large-scale production of functional human lysozyme from marker-free transgenic cloned cows.Scientific reports · 2016Article
- Characterization of promoter of the tuberculosis-resistant gene intracellular pathogen resistance 1.Immunologic research · 2016Article
- Effects of Genetically Modified Milk Containing Human Beta-Defensin-3 on Gastrointestinal Health of Mice.PloS one · 2016Article
- Development of Marker-Free Insect-Resistant Indica Rice byFrontiers in plant science · 2016Article
- Production of transgenic cattle highly expressing human serum albumin in milk by phiC31 integrase-mediated gene delivery.Transgenic research · 2015Article
- Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk.Scientific reports · 2015Article
- Exogenous enzymes upgrade transgenesis and genetic engineering of farm animals.Cellular and molecular life sciences : CMLS · 2015Review
- Induced pluripotent stem cells: Mechanisms, achievements and perspectives in farm animals.World journal of stem cells · 2015Review
- New applications for phage integrases.Journal of molecular biology · 2014Review
- Improved site-specific recombinase-based method to produce selectable marker- and vector-backbone-free transgenic cells.Scientific reports · 2014Article
- Pseudo attP sites in favor of transgene integration and expression in cultured porcine cells identified by Streptomyces phage phiC31 integrase.BMC molecular biology · 2013Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Antibiotic selectable marker genes have been widely used to generate transgenic animals. Once transgenic animals have been obtained, the selectable marker is no longer necessary but raises public concerns regarding biological safety. The aim of this study was to prepare competent antibiotic selectable marker free transgenic cells for somatic cell nuclear transfer (SCNT). PhiC31 intergrase was used to insert a transgene cassette into a "safe harbor" in the bovine genome. Then, Cre recombinase was employed to excise the selectable marker under the monitoring of a fluorescent double reporter. By visually tracking the phenotypic switch from red to green fluorescence, antibiotic selectable marker free cells were easily detected and sorted by fluorescence-activated cell sorting. For safety, we used phiC31 mRNA and cell-permeant Cre protein in this study. When used as donor nuclei for SCNT, these safe harbor integrated marker-free transgenic cells supported a similar developmental competence of SCNT embryos compared with that of non-transgenic cells. After embryo transfer, antibiotic selectable marker free transgenic cattle were generated and anti-bacterial recombinant human β-defensin-3 in milk was detected during their lactation period. Thus, this approach offers a rapid and safe alternative to produce antibiotic selectable marker free transgenic farm animals, thereby making it a valuable tool to promote the healthy development and welfare of transgenic farm animals.
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