Final Activity Report Summary - TEAMOHOLIC (Team on Advanced Transgenic Animal Technologies)
The project successfully achieved its overall goal to strengthen a newly established research team in Hungary on animal biotechnology and to fully integrate it into the European Research Area (ERA). The project created the only Nuclear replacement (NR, 'cloning') team in the Central-European region which successfully developed and demonstrated its world class technological excellence in mouse and rabbit NR. This platform provide the establishment of new transgenic laboratory animal models for medical purposes which will reveal the correlation between the genetic code and the biological functions and potentially clarify the genetic base of the most important diseases affecting the European population.
The objectives in TEAMOHOLIC were to produce the first targeted genetic rabbit, validate NR as a technical platform to produce transgenic mouse models and to develop an improved system for banking and distributing the newly generated models.
The 'Genetic reprogramming' team was greatly strengthened by the TEAMOHOLIC project. Mostly researchers returning from abroad and international researchers have been selected representing a broad spectrum of expertise. Professional trainings helped their personnel development. The success in the integration into the ERA is demonstrated by participation in Sixth Framework Programme (FP6) projects, including a STREP on mouse and rat cloning (MedRat as co-ordinator), a project on ethical, safety and legal aspects of interspecies NR (Chimbrids) and a MC RTN project on biotechnology for new medical animal models (Clonet as coordinator). The uniqueness of the team in Hungary both in its topic (cloning) and establishment (returning scientists after 'brain drain') created a high profile for public perception, media and science politics. The team educated the general public about animal biotechnology via lectures and numerous media events. The scientific community has been informed by the publication of about 30 papers, book chapters and conference proceedings.
In mouse piezo-enucleation and 'zona-free' micromanipulation methods have been compared. Parthenogenetic activation, in vitro culture, sperm, oocyte and embryo cryopreservation and co-transfer protocols were developed further. NR embryos were used to generate new ES cell lines analysed for gene expression patterns showing that they are very similar to the regular ES cells, important information for human cell therapy. The gene targeting work in ES and somatic cells resulted in new knock-out ES cell lines. The first live NR mouse in Hungary was born in November 2006, cloned from cumulus cells followed by another 7 female mice which gave birth to over one hundred healthy pups. From Dll1+/lacZ (Dll1tm1Gos) knock-out donor cumulus cells several cloned mice were born and the breeding of the surviving knock-out cloned female founder is expected to result in sufficient number of animals for full geno- and phenotyping by end of 2008, thus completing all the mouse deliverables. The results are expected to provide novel information about the trans-generational effect of NR, a major issue for the safety of the technology.
In rabbit oocyte collection, in vitro maturation, activation, in vitro culture, sperm and embryo cryopreservation, Trichostatin A treatment (TSA) and pregnancy maintenance have been perfected. Two NR systems were compared, resulting in the birth of somatic cell cloned progeny in May 2007. A large number of live progeny (13) have been born, and 4 of them survived to adulthood and reproduced successfully, thus creating the second largest population of cloned rabbits of the world (after the pioneering INRA, France). The ultimate goal, the production of gene-targeted rabbit have been attempted, however, no live progeny have been obtained - this was not achieved yet by any of the teams working on rabbit NR around the world, neither - showing the important species differences and particular difficulties in rabbit NR.
The objectives in TEAMOHOLIC were to produce the first targeted genetic rabbit, validate NR as a technical platform to produce transgenic mouse models and to develop an improved system for banking and distributing the newly generated models.
The 'Genetic reprogramming' team was greatly strengthened by the TEAMOHOLIC project. Mostly researchers returning from abroad and international researchers have been selected representing a broad spectrum of expertise. Professional trainings helped their personnel development. The success in the integration into the ERA is demonstrated by participation in Sixth Framework Programme (FP6) projects, including a STREP on mouse and rat cloning (MedRat as co-ordinator), a project on ethical, safety and legal aspects of interspecies NR (Chimbrids) and a MC RTN project on biotechnology for new medical animal models (Clonet as coordinator). The uniqueness of the team in Hungary both in its topic (cloning) and establishment (returning scientists after 'brain drain') created a high profile for public perception, media and science politics. The team educated the general public about animal biotechnology via lectures and numerous media events. The scientific community has been informed by the publication of about 30 papers, book chapters and conference proceedings.
In mouse piezo-enucleation and 'zona-free' micromanipulation methods have been compared. Parthenogenetic activation, in vitro culture, sperm, oocyte and embryo cryopreservation and co-transfer protocols were developed further. NR embryos were used to generate new ES cell lines analysed for gene expression patterns showing that they are very similar to the regular ES cells, important information for human cell therapy. The gene targeting work in ES and somatic cells resulted in new knock-out ES cell lines. The first live NR mouse in Hungary was born in November 2006, cloned from cumulus cells followed by another 7 female mice which gave birth to over one hundred healthy pups. From Dll1+/lacZ (Dll1tm1Gos) knock-out donor cumulus cells several cloned mice were born and the breeding of the surviving knock-out cloned female founder is expected to result in sufficient number of animals for full geno- and phenotyping by end of 2008, thus completing all the mouse deliverables. The results are expected to provide novel information about the trans-generational effect of NR, a major issue for the safety of the technology.
In rabbit oocyte collection, in vitro maturation, activation, in vitro culture, sperm and embryo cryopreservation, Trichostatin A treatment (TSA) and pregnancy maintenance have been perfected. Two NR systems were compared, resulting in the birth of somatic cell cloned progeny in May 2007. A large number of live progeny (13) have been born, and 4 of them survived to adulthood and reproduced successfully, thus creating the second largest population of cloned rabbits of the world (after the pioneering INRA, France). The ultimate goal, the production of gene-targeted rabbit have been attempted, however, no live progeny have been obtained - this was not achieved yet by any of the teams working on rabbit NR around the world, neither - showing the important species differences and particular difficulties in rabbit NR.