ArticleTransgenic research2005
Increased transgene integration efficiency upon microinjection of DNA into both pronuclei of rabbit embryos.
Article in Transgenic research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 55 citations in OpenAlex.
- Perspectives in Genome-Editing Techniques for Livestock.Animals : an open access journal from MDPI · 2023Review
- Review
- Investigation of hFVIII production in mammary glands of transgenic mice.Monoclonal antibodies in immunodiagnosis and immunotherapy · 2014Article
- Generation of RAG 1- and 2-deficient rabbits by embryo microinjection of TALENs.Cell research · 2013Article
- Use of transgenic animals in biotechnology: prospects and problems.Acta naturae · 2013Article
- Functional factor VIII made with von Willebrand factor at high levels in transgenic milk.Journal of thrombosis and haemostasis : JTH · 2011Article
- Generation and characterization of a GFP transgenic rat line for embryological research.Transgenic research · 2008Article
- Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits.Transgenic research · 2007Article
- Establishment and characterization of CAG/EGFP transgenic rabbit line.Transgenic research · 2007Article
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Transgenic rabbits provide a useful biological model for the study of the regulation of mammalian genes. However, transgene integration efficiency has generally been low. Here we present a first attempt to increase the integration rate of exogenous DNA into the rabbit genome, using a double pronuclei microinjection method. Pronuclear stage rabbit embryos were recovered from superovulated NZW females, 19-20 h after hCG injection. About 5 microg/mL of exogenous DNA solution was microinjected either into one pronucleus (single microinjection, SM) or into both pronuclei (double microinjected, DM). The transgene consisted of a 2.5 kb murine whey acidic protein promoter (mWAP), 7.2 kb cDNA of the human clotting factor VIII (hFVIII), and 4.6 kb that of 3' flanking sequences of the mWAP gene. The in vitro survival of DM embryos to the blastocyst stage was lower than that of SM embryos (68 vs. 89%). Similar results were obtained using EGFP as a control gene construct. However, there was no difference in the percentage of embryos that developed into live offspring using DM (25%) vs. SM (26%). The integration frequency of mWAP-hFVIII into the genome of transgenic rabbits was 3.3% (1/30) upon SM and 8.1% (4/49) at DM (p < 0.05). All founders transmitted the transgene to their offspring in a Mendelian fashion. The SM founder female secreted 87.4 microg/mL rhFVIII in milk, with an activity of 0.594 IU/mL. The DM founder female produced 118 microg/mL rhFVIII, with activity values of 18 IU/ mL. This is the first report of transgenic rabbit production using a double microinjection technique. Our preliminary results suggest that this method can increase the efficiency of production of transgenic rabbit founders, giving a higher integration rate than single microinjection.
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