Sigma-Aldrich Technology Workshop – 24th March
Sigma-Aldrich are pleased to offer this half-day, fully-sponsored CRISPR Technology workshop on 24th March (morning). Registration for this workshop is free. You are automatically invited to this event if you have already registered for the CRISPR Symposium on 23rd March.
Sigma-Aldrich has an extensive long-term experience in developing technologies for Targeted Genome Editing. Since 2008 they have offered validated ZFN-based kits and are now leading the development of new CRISPR/Cas9 associated products and services.
During this Day 2 of the CRISPR 2015 Oxford Symposium, scientist from Sigma-Aldrich as well as specially invited UK-based researchers will present a series of CRISPR talks that will further enhance your knowledge on Targeted Genome Editing in general and on the CRISPR/Cas9 technology in particular.
Provisional Agenda
Monday 23rd March
19.00: Sigma-Aldrich Sponsored Dinner (Venue TBC)
Tuesday 24th March
08.00: Registration
08.30: Welcome
08.45: Introduction to CRISPR Core Partnership Programme
09.00: Genome Editing with CRISPR/Cas9
Dr Erika Holroyd, Sigma-Aldrich Product Manager
Abstract: CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break at a desired genomic location. The cell then activates endogenous DNA repair processes, either non-homologous end joining or homology-directed repair to heal the targeted double-strand break. This talk will focus on the CRISPR/Cas9 system for genome engineering. Topics will include (i) how to utilize the system for gene knockout or targeted integration, (ii) design, efficiency and specificity of CRISPR, and (iII) how to introduce and validate this nuclease in your model system.
09.30: Coffee break
10.00: Crispr gene knockout and knockin: dissection of metabolic pathways affecting energy metabolism, blood flow and neuroinflammation (Co-authors: James Scott, Sophie Piper, Naveenan Navaratnam, David Carling, James Leiper and Zoe Webster)
Professor James Scott, PhD, Imperial College London, UK
Abstract: The last time we used conventional approaches to generate knockout mice it took us two years to get pups. Routinely using custom Crispr we now create knockout and knockin mice or rat pups in six weeks and cells in three weeks. We have been able to approach the dissection of an entire pro-inflammatory metabolic pathway engendered by Western diet feeding using Crispr. I will present results accordingly of generation multiple knockout mice and rats using custom Crisprs, efficiency data using different strategies, and frequency of off-target effects. Outcomes will be presented in relation to the role of fatty acid metabolic pathways and the role of Tlrs, Myd88 and Eif2ak2 in neuroinflammation and the metabolic induction of App and Bace1 induction, and the possible role of these pathways in Alzheimer’s disease pathogenesis.
10.45: Practical Considerations for Successful Cell Line Engineering
Dr Erika Holroyd, Sigma-Aldrich Product Manager
Abstract: In addition to identifying an active CRISPR reagent to generate your model cell line, creating a genetically modified cell line requires a deep understanding of genetic engineering constraints. Understanding your target cell line and gene of interest play a crucial role in the success of a project. Initial design is critical since the subsequent technical commitment is large, and issues such as isolation of low frequency modifications can make conceptually simple engineering goals unsuccessful. This talk will address some of the practical considerations for successful cell line engineering including (i) cell type, (ii) gene of interest, (iii) engineering method and (iv) screening methods for your modification of interest.
11.15: Examining Radiosensitivity in Human cells: Construction of a cell line library defective in the DNA double strand break repair using CRISPRs
Dr Christopher Parris, PhD, Brunel University, UK
Abstract: TBA
12.00: CRISPR Q&A Session
Facilitated by Dr Erika Holroyd, Sigma-Aldrich Product Manager
12.30: Closing Remarks and Future Outlook
Dr Ed Roullard, Sigma-Aldrich VP Research Markets, EMEA
13.00: Buffet Lunch
