CRM scientists have been able to make precise genetic changes to mouse and human neuronal stem cells for the first time. This opens up the possibility for the study of the genetic changes seen in glioblastoma, the most common form of brain cancer.
Dr Steve Pollard and his team used CRISPR/Cas technology to modify the genes of neuronal stem cells in several different ways, including engineering two mutations found in glioblastoma (deletion of the tumour suppressor gene TP53 and a point mutation H3F3A). This will allow the more detailed study of the precise effects of different genes and mutations in neuronal stem cells.
Genome editing is the process by which the cells blueprint or DNA is cut using a nuclease enzyme to allow modification of the DNA sequence or insertion of a new gene. CRISPR/Cas9 technology uses a the transcription enzyme Cas9 and a molecular guide (CRISPR) to precisely target the cut.
CRISPR/Cas technology has caused a scientific revolution in genetic modification. Dr Pollard said,
“CRISPR has completely transformed the landscape for how we study genetics. It’s a particularly useful tool that’s opening up the human genome for us to be able to do what we want genetically.”
"Most groups have been using CRISPR to make these random changes to genes, which is its simplest use, but we focussed on more sophisticated and precise manipulations, in both mouse and human cells, and were able to make larger-scale genetic changes."
As well as improving the understanding of how neuronal stem cells develop and behave in disease these techniques may now allow scientists to develop new treatments
"The obvious next step is finding out if we can use the same technology in cells taken from patients’ brain tumours, we could make target molecules fluorescent in these patient cells and see which drugs are important in destroying that fluorescence."
Image: Neural stem cells (mouse) with white cells in the process of cell division.
This paper is published in the journal Development.
This work was supported by funding from Cancer Research UK, the Science Without Borders Program (Coordenação de Aperfeiçoamento de Pessoal de Nı́vel Superior, Governo Dilma Rousseff, Brazil) and Fundación Ramón Areces (Spain). Dr Steve Pollard is supported by a Cancer Research UK Senior Research Fellowship (A19778).