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Project Description

If one fact is true: Biology proved itself to be more complex than we could ever have imagined. This especially holds true for fields like cancer research and cell reprogramming. Up to several years ago the main approach for investigating gene signaling cascades was to focus on single genes and underestimating crosstalk in between pathways. Investigating on those crosstalks was a difficult, time consuming work, that is up to date a challenge for scientists all around the world. High throughput methods like whole transcriptome analysis gave rise to complete new approaches and ideas. One major flaw of these methods is, that they are only descriptive and do not allow to interfere with those systems.

Here, we demonstrate a system, based on a protein-RNA-DNA interaction that allows multiple gene regulation simultaneously with minimal effort. Cas9, a nuclease, derived from a bacterial immunity system, is able to bind to multiple RNA specified targets. The FreiGEM 2013 team aims to develop from this protein a new class of DNA binding proteins by mutating the nuclease and by fusing it to effector domains. The aim was not only to modify gene expression, but also to make this process inducible in a spatiotemporal manner. This gives rise to application in medical and fundamental research.

For a proof-of-principle work we demonstrate function in human cell lines and evaluated those results by theoretical modeling. Furthermore, we were able to manipulate endogenous targets in the well-studied model organisms Danio rerio and Arabidopsis thaliana, by targeting factors involved into stem cell reprogramming stem cells were induced and endogenous reporters, respectively.