Optogenetic platforms and optical regulation of Oct4: light-controlled reprogramming

Optogenetics is an emerging synthetic biology method and experimental paradigm for manipulating and studying biological systems in health and disease. At its core, optogenetics utilizes a broad range of light-sensitive proteins as molecular switches that are activated with high temporal precision (e.g. during specific stages of development or in specific phases of an industrial process) and spatial precision (e.g. at specific cellular locations or in specific tissues). The overall goal of this project is to build new and generally applicable capacities in optogenetics research and to apply these capacities to gain more precise control over human cell reprogramming. First, we will obtain genetic parts coding for light-sensitive domains (LSDs) and build a modular genetic engineering platform for generating libraries of optogenetic regulators that can be applied to a myriad of target proteins and cellular processes. Second, we will develop new light delivery hardware with high flexibility for optogenetic experiments and beyond. Finally, we will demonstrate the combined value of the genetic parts and the new devices to the biology of stem cells by demonstrating for the first time optical control of a transcriptional master regulator and of reprogramming. Precise control of the transcription factor (TF) octamer-binding TF 4 (Oct4) will provide a new tool to study cellular fate determination and offer new avenues in the industrial generation of human induced pluripotent stem cells (iPSCs). This project benefits from the skills and experience that the fellow brings from a world-leading laboratory in Europe to Australia and from new synergies between the host organisation and CSIRO. The proposed research aligns with the Application Domains “Health” and “Foundation Technologies” and involves capacity building in the Science Domains “Engineering Novel Biological Components” and “Assembling Novel Biosystems”