OxTalks will soon move to the new Halo platform and will become 'Oxford Events.' There will be a need for an OxTalks freeze. This was previously planned for Friday 14th November – a new date will be shared as soon as it is available (full details will be available on the Staff Gateway).
In the meantime, the OxTalks site will remain active and events will continue to be published.
If staff have any questions about the Oxford Events launch, please contact halo@digital.ox.ac.uk
Recent adaptation of the CRISPR/Cas9 bacterial system to facilitate manipulation of mammalian genomes has provided a real breakthrough for genome editing applications. Development of whole-genome CRISPR libraries with the aim of generating gene knockouts for every single coding sequence has allowed forward genetic screening in mammalian cells with unprecedented efficiency and versatility. CRISPR/Cas9 approaches, however, rely on phenotypes associated with loss-of-function mutations. Single-nucleotide variations (SNVs), on the other hand, have the potential to uncover not only loss-of-function phenotypes (by generating nonsense mutations, for example) but also gain-of-function phenotypes through missense mutations. In addition, they produce valuable information regarding functionally important domains of the affected gene product, as SNVs causative of a particular phenotype tend to cluster around specific regions of the amino acid sequence of the encoded protein. SNV-based approaches in mammalian cells, however, have been hindered by the diploid nature of their genomes, a fact that complicates the establishment of straightforward genotype-to-phenotype correlations. In this talk I will discuss how we apply CRISPR/Cas9 genetic screening to further understand the DNA-damage response in mouse embryonic stem cells (mESCs), and I will also introduce the use of haploid mESCs to perform SNV-based forward genetic screens.