| Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Outline of Final Research Achievements |
Our purpose of this study is to produce genetically modified piglets that are used for xenotransplantation. α-GalT is known to synthesize α-Gal epitope (α-Gal), which is a causative for hyperacute rejection of pig grafts. We first disrupted the α-GalT gene in porcine cells using a recently developed technology that allows efficient ablation of a target gene, and finally obtained a cell line called CRISPR-2-1 (2-1). DAF and TM are also known as key factors suppressing graft rejection. In order to deliver genes for these two factors into 2-1, we developed a novel system that does not require drug selection and also employed a piggyBac-based system that permits introduction of multiple transgenes. We are now constructing 2-1-based new cell lines that are able to express both DAF and TM in the absence of α-Gal. The resulting cell line will be used for SCNT as donors to produce cloned piglets capable of expressing DAF and TM, but unable to express α-Gal.
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