A5018214540- Cancer, Hypoxia, and Metabolism
- Acute Lymphoblastic Leukemia research
- Acute Myeloid Leukemia Research
- CRISPR and Genetic Engineering
- RNA modifications and cancer
- Epigenetics and DNA Methylation
- RNA Research and Splicing
- Protein Degradation and Inhibitors
- Lung Cancer Treatments and Mutations
- PARP inhibition in cancer therapy
- CAR-T cell therapy research
- Neuroblastoma Research and Treatments
- Genomics and Chromatin Dynamics
- Histone Deacetylase Inhibitors Research
- RNA and protein synthesis mechanisms
- RNA Interference and Gene Delivery
- Mitochondrial Function and Pathology
- Pluripotent Stem Cells Research
- Ubiquitin and proteasome pathways
- Immune Cell Function and Interaction
- Cancer Genomics and Diagnostics
- Cancer therapeutics and mechanisms
- Hemoglobinopathies and Related Disorders
- Virus-based gene therapy research
- Inflammasome and immune disorders
St. Jude Children's Research Hospital
2017-2025
Memphis Health Center
2024
Ducks Unlimited
2023
Oak Ridge National Laboratory
2023
Institute of Molecular and Cell Biology
2018-2021
Institute of Hematology & Blood Diseases Hospital
2019
Washington University in St. Louis
2013-2018
The University of Texas Southwestern Medical Center
2008-2013
Sigma Research (United States)
2011
Southwestern Medical Center
2008
The T7 endonuclease 1 (T7E1) mismatch detection assay is a widely used method for evaluating the activity of site-specific nucleases, such as clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system. To determine accuracy and sensitivity this assay, we compared editing estimates derived by T7E1 with that targeted next-generation sequencing (NGS) in pools edited mammalian cells. Here, report nuclease determined most often do not accurately reflect observed Editing...
New insights into norovirus entry There's no escaping when you have it—the symptoms from this gastroenteritis-causing virus, though brief, are often debilitating. Preventing infections will rely on improving our understanding of how enters host cells. Orchard et al. show that the murine (MNoV) cells requires a protein called CD300lf. In cell culture, mouse needed to express CD300lf in order for MNoV binding, entry, and replication occur. Deleting gene encoding mice protected them against...
DNMT3A deletion preserves CAR T cell functionality during prolonged stimulation.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for disease 2019 (COVID-19), continues to cause substantial morbidity and mortality in ongoing global pandemic. Understanding fundamental mechanisms that govern innate immune inflammatory responses during SARS-CoV-2 infection is critical developing effective therapeutic strategies. Whereas interferon (IFN)–based therapies are generally expected be beneficial viral infection, clinical trials COVID-19 have...
Key Points Cas9 editing of the γ-globin gene promoters in hematopoietic stem cells (HSCs) increases red cell HbF by ≤40%. No deleterious effects on hematopoiesis or off-target mutations were detected 16 weeks after xenotransplantation edited HSCs.
CRISPR-Cas9 technology allows the creation of user-defined genomic modifications in cells and whole organisms. However, quantifying editing rates pools or identifying correctly edited clones is tedious. Targeted next-generation sequencing provides a high-throughput platform for optimizing reagents modified clones, but large amount data produced can be difficult to analyze. Here, we present CRIS.py, simple highly versatile python-based program which concurrently analyzes both knock-out...
Significance Primary template-directed amplification (PTA) is a major improvement in whole genome amplification, which required to study intratissue cellular evolution. As presented the manuscript, PTA produces significantly improved and reproducible sequencing coverage variant detection from single of cell. Applications measuring genetic diversity cells with manuscript include examining acquisition changes during normal development aging, consequences specific perturbations such as editing,...
CD8+ cytotoxic T cells (CTLs) orchestrate antitumour immunity and exhibit inherent heterogeneity1,2, with precursor exhausted (Tpex) but not terminally (Tex) capable of responding to existing immunotherapies3-7. The gene regulatory network that underlies CTL differentiation whether Tex cell responses can be functionally reinvigorated are incompletely understood. Here we systematically mapped causal networks using single-cell CRISPR screens in vivo discovered checkpoints for differentiation....
NLRs constitute a large, highly conserved family of cytosolic pattern recognition receptors that are central to health and disease, making them key therapeutic targets. NLRC5 is an enigmatic NLR with mutations associated inflammatory infectious diseases, but little known about its function as innate immune sensor cell death regulator. Therefore, we screened for NLRC5's role in response infections, PAMPs, DAMPs, cytokines. We identified acts drive death, PANoptosis, specific ligands,...
Innate immunity provides the first line of defense through multiple mechanisms, including pyrogen production and cell death. While elevated body temperature during infection is beneficial to clear pathogens, heat stress (HS) can lead inflammation pathology. Links between pathogen exposure, HS, cytokine release, have been observed, but fundamental innate immune mechanisms driving pathology exposure HS remain unclear. Here, we use genetic approaches elucidate pathways in or LPS models. Our...
During development, morphogens pattern tissues by instructing cell fate across long distances. Directly visualizing morphogen transport in situ has been inaccessible, so the molecular mechanisms ensuring successful delivery remain unclear. To tackle this longstanding problem, we developed a mouse model for compromised sonic hedgehog (SHH) and discovered that endocytic recycling promotes SHH loading into signaling filopodia called cytonemes. We optimized methods to preserve vivo cytonemes...
Zinc finger nucleases (ZFNs) have been used successfully to create genome-specific double-strand breaks and thereby stimulate gene targeting by several thousand fold. ZFNs are chimeric proteins composed of a specific DNA-binding domain linked non-specific DNA-cleavage domain. By changing key residues in the recognition helix domain, one can alter ZFN binding specificity change sequence which pair is being targeted. For these other reasons, pursued as reagents for genome modification,...
Z-DNA-binding protein 1 (ZBP1) is an innate immune sensor of nucleic acids that regulates host defense responses and development. ZBP1 activation triggers inflammation pyroptosis, necroptosis, apoptosis (PANoptosis) by activating receptor-interacting Ser/Thr kinase 3 (RIPK3), caspase-8, the NLRP3 inflammasome. unique among sensors because its N-terminal Zα1 Zα2 domains, which bind to in Z-conformation. However, specific role these Zα domains orchestrating subsequent cell death not clear....
Abstract In addition to mutations in genes, aberrant enhancer element activity at non-coding regions of the genome is a key driver tumorigenesis. Here, we perform epigenomic profiling cohort more than forty genetically diverse human colorectal cancer (CRC) specimens. Using normal colonic crypt epithelium as comparator, identify enhancers with recurrently gained or lost across CRC Of highly activated CRC, most are constituents super enhancers, occupied by AP-1 and cohesin complex members,...
Abstract Aggressive cancers often have activating mutations in growth-controlling oncogenes and inactivating tumor-suppressor genes. In neuroblastoma, amplification of the MYCN oncogene inactivation ATRX gene correlate with high-risk disease poor prognosis. Here we show that are mutually exclusive across all ages stages neuroblastoma. Using human cell lines mouse models, found elevated expression incompatible. Elevated levels promote metabolic reprogramming, mitochondrial dysfunction,...
Survival in high-risk pediatric neuroblastoma has remained around 50% for the last 20 years, with immunotherapies and targeted therapies having had minimal impact. Here, we identify small molecule CX-5461 as selectively cytotoxic to synergistic low picomolar concentrations of topoisomerase I inhibitors improving survival vivo orthotopic patient-derived xenograft mouse models. recently progressed through phase clinical trial a first-in-human inhibitor RNA-POL I. However, also use...