B: Novel transgenic strategies

B: Novel transgenic strategies

Research Area B is developing and characterising genetically multi-modified donor pigs to overcome rejection mechanisms, functional incompatibilities, and zoonotic risks. B1 and B2 has employed gene stacking and combineering technology to generate 5´tg pigs carrying hCD46, hCD55, hCD59, hHO1 and hA20. A strategy for dynamic LEA29Y expression in response to rejection has been devised. Moreover, 4´ko pigs that lack SLA class I, GGTA1, CMAH and B4GALNT2 have also been produced. Combination of these genotypes will provide multi-modified pigs for transplantation of xeno-hearts and heart valves. B3 focuses on optimised donor pigs for xenogeneic islet transplantation. Pigs that lack GGTA1 and CCL2 and express LEA29Y and hPD-L1 (2´ko, 2´tg) have been produced, and a hCD47 transgene will be added to overcome cellular rejection by macrophages. Several pig lines with reporter genes are available for studies of beta-cell maturation, function and survival after transplantation. Single-cell RNA sequencing will be employed to inform and improve methods of maturing neonatal porcine islets (NPIs). Both B projects use donor animals with a PERV-C free genetic background.