The introduction of CRISPR/Cas9 technology has taken the functional genomics field by storm, allowing genomic alterations with high degree of precision and throughput, thanks to the availability of genome-wide libraries.
These libraries are either formatted as “pooled”, where guide RNAs, usually packed in lentiviral vectors, are introduced to the entire cell population and selected, or “arrayed”, where the single guide RNAs are introduced into individual populations in microtiter plates allowing the manipulation of a single gene per well. Both approaches present considerable challenges at various levels: in the case of arrayed screen, one of the bottlenecks is the simultaneous and efficient delivery of both the Cas9 protein and the guide RNAs nucleic acid into the cells.
Dr. Barsacchi presents the solutions his team has developed to tackle delivery in the context of image based phenotypic assays, to be deployed in future genome-wide campaigns. He focuses on the development and validation of procedures of transient transfection either using complexes of recombinant CRISPR-Cas9 and guide RNA, or adopting an uncoupled approach where the Cas9 is delivered using the BACMAM system, followed by guide RNA transfection.
Presenter
Rico Barsacchi received his PhD from the Dibit, Scientific Institute San Raffaele in Milan, Italy, with a thesis on modulations of intracellular signaling by nitric oxide in the group of Jacopo Meldolesi. He then moved to the Max Planck Institute of Molecular and Cell Biology in Dresden, as a post doc in Elly Tanaka’s group, working on mechanisms of muscle regeneration. In 2006, he joined the Technology Development Study Core Facility of the same institute where he is still currently working, under the direction of Marc Bickle.