In the last two years I have been working on genome scale genetic screens using the CRISPR-Cas9 gene editing tool. The aim of these screens is to identify genes which confer resistance to chemotherapy drugs in embryonic stem cells and to oxidative stress agents in dopamine neurons. In the last 9 years I have mostly worked on understanding midbrain dopamine neuron specification and differentiation. More specifically, I studied the roles of transcription factors Foxa1 and Foxa2 in dopamine neuron development through ChIP-Seq and RNA-Seq experiments using in vitro and in vivo mouse models of dopamine neurons.
One of the most interesting and challenging projects I was recently involved in was to produce the first genome wide, arrayed guide RNA screening libraries for CRISPR-Cas9. These libraries have been designed to cover 17166 human and 20430 mouse genes. Two gRNAs are designed per gene giving a total of 34332 gRNAs and 40860 gRNAs for human and mouse genomes, respectively.
It is amazing to be able to undertake such large scale projects here at WTSI, and I am looking forward to the next challenges ahead.
Genome-wide characterisation of Foxa1 binding sites reveals several mechanisms for regulating neuronal differentiation in midbrain dopamine cells.
Development (Cambridge, England) 2015;142;7;1315-24
A conditional piggyBac transposition system for genetic screening in mice identifies oncogenic networks in pancreatic cancer.
Nature genetics 2015;47;1;47-56
Foxa1 and foxa2 are required for the maintenance of dopaminergic properties in ventral midbrain neurons at late embryonic stages.
The Journal of neuroscience : the official journal of the Society for Neuroscience 2013;33;18;8022-34
Genome-wide characterization of Foxa2 targets reveals upregulation of floor plate genes and repression of ventrolateral genes in midbrain dopaminergic progenitors.
Development (Cambridge, England) 2012;139;14;2625-34
Foxa1 and Foxa2 positively and negatively regulate Shh signalling to specify ventral midbrain progenitor identity.
Mechanisms of development 2011;128;1-2;90-103
Foxa1 and Foxa2 function both upstream of and cooperatively with Lmx1a and Lmx1b in a feedforward loop promoting mesodiencephalic dopaminergic neuron development.
Developmental biology 2009;333;2;386-96