Engineering of Human Stem Cells Using Lentiviruses

Very long, linear DNA molecules carry the inheritable and instructive information of an organisms’ genome that is at the foundation of life. To be packaged and protected, genomes are organized into chromatin. This process is not static but highly dynamic constituting a fundamental regulatory system. Different proteins of the linker histone family are crucial for organizing DNA into chromatin. Various, often sporadic findings point to changes in linker histone expression and/ or changes in their composition (aka mutation) to be linked to various diseases including diabetes, cardiovascular disease and cancer. We have set up different experimental systems to thoroughly analyze the role of linker histones in cell differentiation, organismal development and tissue homeostasis. In this VSRP project, we want to adopt lentiviruses for perturbing the expression of linker histones in human embryonic stem cells. We will engineer knockdown (using siRNA) and overexpression schemes to analyze how changes in the levels of linker histones affect cell pluripotency and stem cell differentiation. The work is embedded into two separate projects that investigate the role of linker histones in myogenesis, and neurodevelopment and besides specific experimentation offers broad insights into different aspects of epigenetics in health and disease.