Elucidation of the role of the anti-apoptotic Bcl-2 family A1 in haematopoietic cell development and tumorgenesis by using RNA interference in teh mouse

• Applicant: Professor Dr. Marco Herold, Ph.D.
• Subject Area: Cell Biology
• Term: from 2008 to 2012
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 73090384

A1, an anti-apoptotic Bcl-2 family member, has been shown to be expressed during embryogenesis and during haematopoietic cell development. Transgenic overexpression of A1 promotes B and T cell survival as well as lymphomagenesis. Due to the fact that mice have four closely linked a1 genes, which are however interspersed with some other unrelated genes, classical knock-out technology does not allow generation of a mouse model lacking all A1 isoforms. Therefore we want to use the recently described micro RNA (miR) like shRNA technology combined with viral transduction and transgenesis methods to knock-down expression of all A1 isoforms in vivo (miR-A1). To study the role of A1 in haematopoietic cells, we will (i) reconstitute lethally irradiated mice with miR-A1 transduced foetal liver-derived haematopoietic stem cells and (ii) stably express a miR-A1 cassette under the control of a haematopoietic tissue specific promoter by lentiviral transgenesis. To examine the role of A1 in pre-B/B cell lymphoma development, we will ablate A1 in foetal liver derived haematopoietic stem cells from Eμ-myc transgenic mice and follow their fate after transfer into irradiated recipient mice. Finally, we will express the miR-A1 cassette under the control of a tetracycline inducible promoter to ablate A1 in malignant Eμ-myc tumours and during embryonic development. In summary, the proposed project will not only allow deeper insights into the functions of A1 in vivo but will also establish novel techniques facilitating the study of many other interesting gene products.

• DFG Programme: Research Fellowships
• International Connection: Australia
• Host: Professor Dr. Andreas Strasser