Investigating the influence of Calreticulin (CALR) mutations on cellular phenotypes
Leitung und Betreuung der Masterarbeit
PD Dr. med. Alexandre Theocharides
Leitung Klinik für Medizinische Onkologie und Hämatologie
Prof. Dr. med. Markus G. Manz
Background: CALR mutations are often found in myeloproliferative neoplasms (MPN), a group of hematopoietic stem cell diseases. Apart from activation of JAK/STAT signaling in mutated cells, little is known about the cellular consequences of mutation of this chaperone protein.
Current status of the project: preliminary data from unbiased proteomics screens have shown significant differences between CALR-mutated and CALR-wildtype cells. The exact relevance of these mutations still needs to be determined.
Aim: determining the influence of inducing CALR mutations in CRISPRed cell lines and primary (transduced) patient material to comprehensively determine phenotypes and their potential relevance for driving MPN disease.
Main techniques: CRISPR, FACS, survival assays, siRNA, Western Blot, RT-qPCR
Potential addition, to be discussed: compare effects of CALR mutations to JAK2 mutations, a second well-known driver mutation of MPN, using primary (transduced) patient material or generated or obtained JAK2-mutated cell lines.
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