A5081061069- CRISPR and Genetic Engineering
- Nuclear Receptors and Signaling
- Genetics and Neurodevelopmental Disorders
- Molecular Biology Techniques and Applications
- Single-cell and spatial transcriptomics
- Autophagy in Disease and Therapy
- Animal Genetics and Reproduction
- 3D Printing in Biomedical Research
- RNA Research and Splicing
- Alzheimer's disease research and treatments
- Genomics and Chromatin Dynamics
- Neurogenesis and neuroplasticity mechanisms
- Endoplasmic Reticulum Stress and Disease
- Cellular transport and secretion
- Neuroinflammation and Neurodegeneration Mechanisms
- Mitochondrial Function and Pathology
- RNA and protein synthesis mechanisms
- Acute Myeloid Leukemia Research
- Parkinson's Disease Mechanisms and Treatments
- Advanced biosensing and bioanalysis techniques
- Neuroscience and Neuropharmacology Research
- Pluripotent Stem Cells Research
- RNA Interference and Gene Delivery
- PARP inhibition in cancer therapy
- Protein Degradation and Inhibitors
UK Dementia Research Institute
2018-2025
University of Cambridge
2018-2025
Babraham Bioscience Technologies (United Kingdom)
2024
Babraham Institute
2023-2024
MRC Toxicology Unit
2023
MRC Mitochondrial Biology Unit
2023
Wellcome Sanger Institute
2014-2019
Medical Research Council
2010-2013
Mutations in the ATM tumor suppressor gene confer hypersensitivity to DNA-damaging chemotherapeutic agents. To explore genetic resistance mechanisms, we performed genome-wide CRISPR-Cas9 screens cells treated with DNA topoisomerase I inhibitor topotecan. Thus, here establish that inactivating terminal components of non-homologous end-joining (NHEJ) machinery or BRCA1-A complex specifically topotecan ATM-deficient cells. We show ATM-mutant poly-(ADP-ribose) polymerase (PARP) olaparib reflects...
Abstract Parkinson’s Disease (PD) is the second most common neurodegenerative disorder. The pathological hallmark of PD loss dopaminergic neurons and presence aggregated α-synuclein, primarily in substantia nigra pars compacta (SNpc) midbrain. However, molecular mechanisms that underlie pathology different cell types not currently understood. Here, we present a single nucleus transcriptome analysis human post-mortem SNpc obtained from 15 sporadic cases 14 Controls. Our dataset comprises ∼84K...
Neurodegenerative diseases like Alzheimer's disease, Parkinson's disease and Huntington's manifest with the neuronal accumulation of toxic proteins. Since autophagy upregulation enhances clearance such proteins ameliorates their toxicities in animal models, we others have sought to re-position/re-profile existing compounds used humans identify those that may induce brain. A key challenge this approach is assess if any hits identified can at concentrations would be seen taking drug for its...
The advent of induced pluripotent stem cell (iPSC)-derived neurons has revolutionized Parkinson's disease (PD) research, but single-cell transcriptomic analysis suggests unresolved cellular heterogeneity within these models. Here, we perform the largest study human iPSC-derived dopaminergic to elucidate gene expression dynamics in response cytotoxic and genetic stressors. We identify multiple neuronal subtypes with transcriptionally distinct profiles differential sensitivity stress,...
Cell functions rely on intracellular transport systems distributing bioactive molecules with high spatiotemporal accuracy. The endoplasmic reticulum (ER) tubular network constitutes a system for delivering luminal solutes, including Ca2+, across the cell periphery. How ER structure enables this nanofluidic is unclear. Here, we show that membrane-localized reticulon 4 (RTN4/Nogo) sufficient to impose neurite outgrowth inhibition in human cortical neurons while acting as an morphoregulator....
The maintained expression of transcription factors throughout the development mesodiencephalic dopaminergic (mDA) neurons suggests multiple roles at various stages in development. Two members forkhead/winged helix factor family, Foxa1 and Foxa2, have been recently shown to an important influence early mDA neurons. Here we present data demonstrating that these genes are also involved later maintenance system. We conditionally removed both postmitotic using dopamine transporter-cre mouse....
Abstract We recently identified the splicing kinase gene SRPK1 as a genetic vulnerability of acute myeloid leukemia (AML). Here, we show that or pharmacological inhibition leads to cell cycle arrest, leukemic differentiation and prolonged survival mice transplanted with MLL -rearranged AML. RNA-seq analysis demonstrates altered isoform levels many genes including several established roles in leukemogenesis such MYB , BRD4 MED24 . focus on its main isoforms have distinct molecular properties...
Assemblies of tau can transit between neurons, seeding aggregation in a prion-like manner. To accomplish this, must cross cell-limiting membranes, process that is poorly understood. Here, we establish assays for the study entry into cytosol as phenomenon distinct from uptake, real time, and at physiological concentrations. The pathway cell type specific and, highly sensitive to cholesterol. Depletion cholesterol transporter Niemann-Pick C1 or extraction membrane renders neurons permissive...
Synapse loss strongly correlates with cognitive decline in Alzheimer's disease (AD), but the underlying mechanisms are poorly understood. Deficient Wnt signaling contributes to synapse dysfunction and AD. Consistently, a variant of LRP6 receptor, (LRP6-Val), reduced signaling, is linked late-onset However, impact LRP6-Val on healthy AD brain has not been examined. Knock-in mice, generated by gene editing, carrying this Lrp6 develop normally. neurons from Lrp6-val mice do respond Wnt7a,...
Primed epiblast stem cells (EpiSCs) can be reverted to a pluripotent embryonic cell (ESC)-like state by expression of single reprogramming factor. We used CRISPR activation perform genome-scale, screen in EpiSCs and identified 142 candidate genes. Our validated total 50 genes, previously not known contribute reprogramming, which we chose Sall1 for further investigation. show that augments mouse fibroblasts these induced are indeed fully including formation chimeric mice. also demonstrate...
Abstract Enhanced expression of the cold‐shock protein RNA binding motif 3 (RBM3) is highly neuroprotective both in vitro and vivo . Whilst upstream signalling pathways leading to RBM3 have been described, precise molecular mechanism cold induction remains elusive. To identify temperature‐dependent modulators RBM3, we performed a genome‐wide CRISPR‐Cas9 knockout screen using RBM3‐reporter human iPSC‐derived neurons. We found that mRNA levels are robustly regulated by several splicing...
Abstract Organelles form membrane contact sites between each other, allowing for the transfer of molecules and signals. Mitochondria-endoplasmic reticulum (ER) (MERCS) are cellular subdomains characterized by close apposition mitochondria ER membranes. They have been implicated in many diseases, including neurodegenerative, metabolic, cardiac diseases. Although MERCS extensively studied, much remains to be explored. To uncover novel regulators MERCS, we conducted a genome-wide, flow...
The transcription factors Foxa1 and Foxa2 promote the specification of midbrain dopaminergic (mDA) neurons floor plate. Whether their role is direct has remained unclear as they also regulate expression Shh, which similar roles. We characterized cis-regulatory network by chromatin immunoprecipitation followed high-throughput sequencing mDA progenitors. This identified 9160 high-quality binding sites associated with 5409 genes, providing mechanistic insights into Foxa2-mediated positive...
CRISPR-Cas9 technology has accelerated biological research becoming routine for many laboratories. It is rapidly replacing conventional gene editing techniques and high utility both genome-wide gene-focussed applications. Here we present the first individually cloned genome wide arrayed sgRNA libraries covering 17,166 human 20,430 mouse genes at a complexity of 34,332 sgRNAs 40,860 genome. For flexibility in generating stable cell lines have been lentivirus backbone containing PiggyBac...
Abstract Accumulation of aggregated and misfolded proteins, leading to endoplasmic reticulum stress activation the unfolded protein response, is a hallmark several neurodegenerative disorders, including Alzheimer’s Parkinson’s disease. Genetic screens are powerful tools that proving invaluable in identifying novel modulators disease associated processes. Here, we performed loss-of-function genetic screen using human druggable genome library, followed by an arrayed-screen validation,...
Abstract Background Microexons, exons that are ≤ 30 nucleotides, a highly conserved and dynamically regulated set of cassette exons. They have key roles in nervous system development function, as evidenced by recent results demonstrating the impact microexons on behaviour cognition. However, often overlooked due to difficulty detecting them using standard RNA-seq aligners. Results Here, we present MicroExonator, novel pipeline for reproducible de novo discovery quantification microexons. We...
Triplications and certain point mutations in the SNCA gene, encoding alpha-synuclein (α-Syn), cause Parkinson's disease (PD). Here, we demonstrate that PD-causing A53T α-Syn mutation elevated expression perturb acetyl-coenzyme A (CoA) p300 biology human neurons CNS of zebrafish mice. This dysregulation is mediated by activation ATP-citrate lyase (ACLY), a key enzyme generates acetyl-CoA cytoplasm, via two mechanisms. First, ACLY activity increases levels, which activate p300. Second, LKB1...