Human Cancer Genetics Programme

Molecular Cytogenetics Unit

Head of Unit:  Sandra Rodríguez
Research highlights
Optimising CRISPR-Cas9 to model cancer aberrations in human stem cells

Efficient methodologies for recreating cancer-associated chromosome translocations are in high demand as tools for investigating how such events initiate cancer. The CRISPRCas9 system has been used to recreate the genetics of these complex rearrangements at native loci while maintaining the architecture and regulatory elements. However, the CRISPR system remains inefficient for gene editing in human stem cells. We have optimised new strategies to enhance the efficiency of CRISPR-mediated translocation induction in human stem cells, including mesenchymal and induced pluripotent stem cells. We found that the generation of targeted translocation is significantly increased when using a combination of ribonucleoprotein complexes (Cas9 protein+sgRNA) and ssODNs. The CRISPRCas9-mediated generation of targeted translocations in human stem cells opens up new avenues to model cancer.

Technological and translational activities

We provide state-of-the-art Molecular Cytogenetic and Genome Editing services. The Unit makes available various techniques, which may provide more sensitive and accurate tools to analyse cancer cells, to research groups ; these techniques include RNA-FISH, chromosome stability studies based on a combined array CGH-FISH approach, or the use of CRISPR libraries to perform high-throughput functional analysis. For gene editing experiments, we have set up a specific PCR-based FISH analysis to detect the genome integration site of small constructs including LV particles. In 2017, we carried out over 1,500 assays for experimental and clinically-oriented projects.