Genome Engineering and Synthetic Biology are revolutionizing Life Sciences. Driven by advances in the CRISPR-toolbox for rapid, cheap, multiplex modification of genomes and breakthroughs in DNA synthesis technologies, the pace of progress enabled by these tools in the last 2 years has been breathtaking.
The 1st Genome Engineering and Synthetic Biology: Tools and Technologies meeting (GESB) in September 2013 was a roaring success. The 2nd edition of GESB once again took place in Ghent, Belgium in January 2016 and brought together some of the most highly regarded academics (collectively having published ˃350 Nature, Cell and Science papers) and companies in the world, with novel technologies in several sessions, with topics focused on:
- CRISPR and Synthetic Biology Tools
- Gene and Genome assembly
- CRISPR for re-engineering cellular systems
- Future CRISPR applications
- Expanding the Genetic Code
In addition to a great scientific and technology program, the conference provided ample opportunities to network during the breaks, poster sessions, the conference dinner and a ‘Meet the Expert’ session with MIT Prof Christopher Voigt.
Tom Henley from Horizon Discovery spoke on Lessons Learned from Hundreds of Individual Genome Engineering Events
The advent of the CRISPR/Cas9 system has revolutionised the field of genome engineering. This technology allows for the rapid generation of mutant human cell lines harbouring diverse genetic modifications. The simplicity of design and ease of use makes the creation of gene knock-outs a relatively straight forward process. However, engineering cell lines with precise genetic changes, such as mutations occurring in human disease or the insertion of DNA to tag genes or create fusions, offers an added level of complexity. Having engineered hundreds of unique and precise mutations in a wide range of cell types, we have begun to learn the fundamental principles that govern the success of generating knock-ins.
I will discuss our experiences and how they have influenced our cell line engineering approach including choices such as the type of donor to use when introducing a mutation, when to incorporate selection to improve efficiency or when to nick the target gene versus inducing a double strand break. As well as design considerations, we have found that in-depth characterisation of an engineering event is essential to ensure the modification has occurred as intended with no unwanted changes on additional alleles. I will also discuss how this validation has its own challenges and requires careful consideration of the techniques used to ensure unpredicted alterations in the genome are revealed.