Improvement of pronuclear injection-based targeted transgenesis (PITT) by iCre mRNA-mediated site-specific recombination
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KeywordsGeneration Vector Transgenic Rate Developmental Arrest Supplementary Movie Donor Vector
We are grateful to K. Yamagata for pcDNA3.1EGFP-poly(A83) (Yamagata et al. 2005), and I. M. Verma for iCre plasmid (Addgene plasmid 12336). We also thank S. Ogiwara and T. Sato for technical assistance. We are grateful to Manju George for critical reading of the manuscript and for English corrections. This work is supported by the Grant-in-Aid for Young Scientists (B) (20700368) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan, by Takeda Science Foundation, by the A-STEP (Adaptable and Seamless Technology Transfer Program through Target-driven R&D) from Japan Science and Technology Agency (JST), and by 2011–2012 Tokai University School of Medicine, Project Research to M.O.
This movie file shows injections of donor vector DNA/iCre mRNA mixture into the zygotes according to the Method 1 (see “Supplementary materials and methods”). These injections are demonstration using BDF2 zygotes (MPG 4624 kb)
This movie file shows injections of donor vector DNA/iCre mRNA mixture into the zygotes according to the Method 2 (see “Supplementary materials and methods”). These injections are demonstration using BDF2 zygotes (MPG 3048 kb)
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