A5010747655- CAR-T cell therapy research
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Immune Cell Function and Interaction
- Advanced biosensing and bioanalysis techniques
- MicroRNA in disease regulation
- Single-cell and spatial transcriptomics
- RNA and protein synthesis mechanisms
- T-cell and B-cell Immunology
- Pluripotent Stem Cells Research
- Axon Guidance and Neuronal Signaling
- Virus-based gene therapy research
- RNA Research and Splicing
- Angiogenesis and VEGF in Cancer
- FOXO transcription factor regulation
- RNA modifications and cancer
- Advanced Memory and Neural Computing
- Viral Infectious Diseases and Gene Expression in Insects
- Biomedical Ethics and Regulation
- Origins and Evolution of Life
- Genomics and Chromatin Dynamics
- Gamma-ray bursts and supernovae
- Neuroscience and Neural Engineering
- Nanowire Synthesis and Applications
- Epigenetics and DNA Methylation
CancerCare Manitoba
2023-2025
University of Manitoba
2023-2025
Research Institute in Oncology and Hematology
2025
University of British Columbia
2020-2024
MRC Weatherall Institute of Molecular Medicine
2016-2024
University of Oxford
2016-2024
Canada's Michael Smith Genome Sciences Centre
2021-2022
Howard Hughes Medical Institute
2016
Massachusetts General Hospital
2016
Abstract Precise, analogue regulation of gene expression is critical for cellular function in mammals. In contrast, widely employed experimental and therapeutic approaches such as knock-in/out strategies are more suitable binary control activity. Here we report on a method precise levels mammalian cells using engineered microRNA response elements (MREs). First, measure the efficacy thousands synthetic MRE variants under an endogenous by high-throughput sequencing. Guided this data, establish...
T cells show tremendous efficacy as cellular therapeutics. However, obtaining primary from human donors is expensive and variable. Pluripotent stem (PSCs) have the potential to provide a renewable source of cells, but differentiating PSCs into hematopoietic progenitors with cell remains an important challenge. Here, we report efficient serum- feeder-free system for cells. This fully defined approach allowed us study impact individual proteins on blood emergence differentiation. Providing...
Abstract Spatial/temporal control of Cas9 guide RNA expression could considerably expand the utility CRISPR-based technologies. Current approaches based on tRNA processing offer a promising strategy but suffer from high background. Here, to address this limitation, we present screening platform which allows simultaneous measurements promoter strength, 5′, and 3′ efficiencies across library variants. This analysis reveals that sequence determinants underlying these activities, while...
Gene editing facilitated by homology-directed repair (HDR) holds great potential for treating monogenetic disorders such as recessive dystrophic epidermolysis bullosa (RDEB). However, low efficiency and variability between loci must be overcome its widespread adoption into personalized therapies. To address these challenges, we developed a highly efficient versatile gene strategy RDEB that incorporates the small molecule inhibitor M3814 to enhance HDR. We focused on three causative COL7A1...
Chimeric Antigen Receptor (CAR) T therapy is a potent treatment for haematological malignancies, but cell exhaustion reduces its efficacy in many patients. Although high CAR transgene levels appear to drive exhaustion, the relationship between expression levels, function, and transcriptional identity yet be mapped at resolution. Here, we harness high-resolution microRNA-based control system precisely modulate assess impact on activation, gene function. By post-transcriptionally modulating...
Abstract The generation of T-cells from stem cells in vitro could provide an alternative source for immunotherapies. T-cell development hematopoietic and progenitor (HSPCs) is tightly regulated through Notch pathway activation by Delta-like (DL) ligands 1 4. Other molecules, such as cell factor (SCF) interleukin (IL)-7, play a supportive role regulating the survival, differentiation, proliferation developing T-cells. Numerous other signaling molecules influence T-lineage vivo, but little...
Abstract Semaphorin-3A (SEMA3A) functions as a chemorepulsive signal during development and can affect T cells by altering their filamentous actin (F-actin) cytoskeleton. The exact extent of these effects on tumour-specific are not completely understood. Here we demonstrate that Neuropilin-1 (NRP1) Plexin-A1 Plexin-A4 upregulated stimulated CD8 + cells, allowing tumour-derived SEMA3A to inhibit cell migration assembly the immunological synapse. Deletion NRP1 in both CD4 enhance T-cell...
A dominant transcription factor mutation disrupting nuclear liquid phase separation causes human immunodeficiency.
Article11 November 2022Open Access Source DataTransparent process Synthetic gene circuits for cell state detection and protein tuning in human pluripotent stem cells Laura Prochazka Institute of Biomedical Engineering (BME), University Toronto, ON, Canada Donnelly Centre Cellular & Biomolecular Research, Contribution: Conceptualization, Data curation, Formal analysis, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Project...
Abstract RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene regulation. Deciphering the prevalence and functional impact this post-transcriptional control layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we describe genome-engineering based evaluation element activity (GenERA), a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 platform in situ high-content analysis RREs. We use...
Ribozyme-catalyzed RNA polymerization is inefficient and error prone. Here we demonstrate that two alternative bases, 2-thio-uridine (s(2)U) 2-thio-ribo-thymidine (s(2)T), improve the rate fidelity of ribozyme catalyzed nucleotide addition as NTP substrates template bases. We also functionality s(2)U s(2)T-containing ribozymes.
Abstract T cells develop from multi-potent hematopoietic progenitors in the thymus and provide adaptive protection against pathogens cancer. However, emergence of human cell-competent blood progenitors, their subsequent specification to lineage, has been challenging capture real time. Here, we leveraged a pluripotent stem cell differentiation system understand transcriptional dynamics fate restriction events that underlie this critical developmental process. Time-resolved single RNA...
ABSTRACT Spatial/temporal control of Cas9 guide RNA expression could considerably expand the utility CRISPR-based technologies. Current approaches based on tRNA processing offer a promising strategy but suffer from high background. Here we developed variant screening platform to identify differential sequence determinants human promoter and activities. Rational design ensuing principles allowed us engineer an improved scaffold that enabled highly specific production Pol-II promoter.
Allogeneic T cell therapies are a highly desirable option to circumvent the cost and complexity of using autologous cells treat diseases. CD8+ can be made from pluripotent stem (PSCs), but deriving CD4+ PSCs remained significant challenge. Using feeder- serum-free conditions, we found that versus commitment controlled by fine-tuning dynamics Notch receptor signaling delivered CD4+CD8+ double positive cells. negatively impacts commitment, its timed removal allows generation clonally-diverse...
T cells develop from hematopoietic progenitors in the thymus and protect against pathogens cancer. However, emergence of human cell-competent blood their subsequent specification to lineage have been challenging capture real time. Here, we leveraged a pluripotent stem cell differentiation system understand transcriptional dynamics fate restriction events that underlie this critical developmental process. Time-resolved single-cell RNA sequencing revealed downregulation multipotent program,...
ABSTRACT T cells are key mediators of the adaptive immune response and show tremendous efficacy as cellular therapeutics. However, obtaining primary from human donors is expensive variable. Pluripotent stem (PSCs) have potential to serve a consistent renewable source cells, but differentiating PSCs into hematopoietic progenitors with cell remains significant challenge. Here, we developed an efficient serum- feeder-free protocol for cells. This defined method allowed us study impact...