February 16, 2015
405 Hilgard Ave, Los Angeles, CA 90095, USA
16-28 February 2015
Wellcome Trust Genome Campus, Hinxton, Cambridge
This lab-based training class will give an all-inclusive review as well as practical lab expertise of the genetic manipulation of mammalian stem cells. Focusing on mouse ES and human iPS cells, genome informatics, recombineering, CRISPR designer nucleases, transposon technology and gene targeting will likely be covered in interactive demonstrations, discussions and lectures by international specialists. Lab work comprises the culture and transfection of mouse ES and human iPS cells, design and construction of gene targeting vectors, as well as the usage of PiggyBac transposons and designer nucleases (CRISPRs) for complex genome engineering and genetic screens. Participants will soon be trained in the informatics as well as practical use of public IKMC (International Knockout Mouse Consortium) mouse knockout resources, for example, generation of mutant mice. Emphasis is put on the preparation/design and successful experimental performance of gene targeting jobs and participants should obtain the skills required to design, build and target their particular genes of interest.
The informatics underlying the visualization and interpretation of gene constructions will likely be presented focusing on the practical design of gene targeting strategies. Pupils may also develop their own CRISPR-mediated gene targeting layouts using internet-based tools and find out the best way to use the public IKMC/IMPC mouse gene targeting/phenotyping resources.
- Recombineering of BACs and Gene Targeting Vectors
Pupils will use recombineering to make changes to BAC transgenes and gene targeting concepts. The theoretical principles underlying both recombineering and reasonable targeting vector layout will soon be shown in lectures and practical exercises.
- Modular Gene Targeting Vectors
Learn to use modular approaches to vector assembly and gene targeting strategies. Students will construct their particular vectors for CRISPR-based gene targeting using Gibson assembly.
- Stem Cell Culturing & Gene Targeting
Pupils will learn feeder-free culture of mouse ES cells and human iPS cells including several conventional transfection techniques (electroporation, nucleofection, lipofection). Transfection, choice, picking, processing and genotyping for gene targeting in ES cells will likely be covered.
- CRISPR Designer Nucleases
A CRISPR mediated homologous recombination experiment will likely be performed in human IPS cells. Practical application of the CRISPR system to undertake in vitro genetic screens may also be investigated.
- Transposon Technology/IPS Cell Creation
Pupils will soon be introduced to the uses of the exceptionally efficient PiggyBac transposon systems for BAC transgenesis and reprogramming for iPS.
To learn more about this conference or to add your international meeting or symposium to our calendar please contact us today.