A5072443724- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- Receptor Mechanisms and Signaling
- RNA and protein synthesis mechanisms
- Protein Kinase Regulation and GTPase Signaling
- RNA modifications and cancer
- Computational Drug Discovery Methods
- Chromatin Remodeling and Cancer
- Antimicrobial Peptides and Activities
- Chromosomal and Genetic Variations
- Chemical Synthesis and Analysis
- Cancer Immunotherapy and Biomarkers
- Immune cells in cancer
- Innovative Microfluidic and Catalytic Techniques Innovation
- Mechanisms of cancer metastasis
- CAR-T cell therapy research
- Cancer Mechanisms and Therapy
- Monoclonal and Polyclonal Antibodies Research
- Virus-based gene therapy research
- Marine Biology and Environmental Chemistry
- Connexins and lens biology
- Plant biochemistry and biosynthesis
- PI3K/AKT/mTOR signaling in cancer
- DNA and Nucleic Acid Chemistry
- Biochemical and Structural Characterization
Pfizer (United States)
2020-2023
Biolog (United States)
2018-2019
Johns Hopkins University
2015
Tufts Medical Center
2013
University of Vermont
1966-1967
The recognition that individual GPCRs can activate multiple signaling pathways has raised the possibility of developing drugs selectively targeting therapeutically relevant ones. This requires tools to determine which G proteins and βarrestins are activated by a given receptor. Here, we present set BRET sensors monitoring activation 12 protein subtypes based on translocation their effectors plasma membrane (EMTA). Unlike most existing detection systems, EMTA does not require modification...
Abstract Close proximity between cytotoxic T lymphocytes and tumour cells is required for effective immunotherapy. However, what controls the spatial distribution of in microenvironment not well understood. Here we couple digital pathology transcriptome analysis on a large ovarian cohort develop machine learning approach to molecularly classify characterize tumour-immune phenotypes. Our study identifies two important hallmarks characterizing cell excluded tumours: 1) loss antigen...
Abstract The success of CRISPR-mediated gene perturbation studies is highly dependent on the quality gRNAs, and several tools have been developed to enable optimal gRNA design. However, these are not all adaptable latest CRISPR modalities or nucleases, nor do they offer comprehensive annotation methods for advanced applications. Here, we present a new ecosystem R packages, called crisprVerse , that enables efficient design multitude technologies. This includes knockout (CRISPRko), activation...
Personalized cancer therapeutics bring directed treatment options to patients based on their tumor's genetic signature. Unfortunately, tumor genomes are remarkably adaptable, and acquired resistance through gene mutation frequently occurs. Identifying mutations that promote within drug-treated patient populations can be cost, resource, time intensive. Accordingly, base editing, enabled by Cas9-deaminase domain fusions, has emerged as a promising approach for rapid, large-scale variant...
Genome-wide loss-of-function screens using the CRISPR/Cas9 system allow efficient discovery of cancer cell vulnerabilities. While several studies have focused on correcting for DNA cleavage toxicity biases associated with copy number alterations, effects sgRNAs co-targeting multiple genomic loci in CRISPR not been discussed. In this work, we analyze essentiality screen data from 391 lines to characterize induced by multi-target sgRNAs. We investigate two types multi-targets: on-targets...
Mutant-specific inhibitors of KRASG12C, such as AMG510 (sotorasib) and MRTX849 (adagrasib), offer the unprecedented opportunity to inhibit KRAS, most frequently mutated heretofore undruggable oncoprotein. While clinical data are still limited, on-target mutations in KRASG12C at position 12 other sites emerging major drivers relapse. We identified additional that impact inhibitor sensitivity through a saturation mutagenesis screen NCI-H358 non–small-cell lung cancer (NSCLC) cell line. also...
Abstract Cas12a is a next-generation gene editing tool that enables multiplexed targeting. Here, we present mouse model constitutively expresses enhanced Acidaminococcus sp. ( enAsCas12a ) linked to an mCherry fluorescent reporter. We demonstrate efficient single and in vitro, using primary transformed cells from mice. further successful vivo editing, normal cancer-prone stem reconstitute the haematopoietic system of wild-type also compact, genome-wide knockout libraries, with four crRNAs...
Target-anchored monovalent degraders are more drug-like than their bivalent counterparts, Proteolysis Targeting Chimeras (PROTACs), while offering greater target specificity control the E3 ligase-anchored degraders, also known as molecular glues. However, discovery has typically been serendipitous, and rules governing identification remain unclear. This study focused on intentional of SMARCA2/A4 using a library based bromodomain-binding ligands. Compound G-6599 emerged lead candidate,...
<title>Abstract</title> Target-anchored monovalent degraders are more drug-like than their bivalent counterparts, Proteolysis Targeting Chimeras (PROTACs), while offering greater target specificity control the E3 ligase-anchored degraders, also known as molecular glues. However, discovery has typically been serendipitous, and rules governing identification remain unclear. This study focused on intentional of SMARCA2/A4 using a library based bromodomain-binding ligands. Compound G-6599...
Ferroptosis is an iron-dependent cell death mechanism characterized by the accumulation of toxic lipid peroxides and membrane rupture. GPX4 (glutathione peroxidase 4) prevents ferroptosis reducing these into alcohols. induction inhibition has emerged as a vulnerability cancer cells, highlighting need to identify regulators that may be exploited therapeutically. Through genome-wide CRISPR activation screens, we SWI/SNF (switch/sucrose non-fermentable) ATPases BRM (SMARCA2) BRG1 (SMARCA4)...
Abstract Motivation: DNA methylation patterns are well known to vary substantially across cell types or tissues. Hence, existing normalization methods may not be optimal if they do take this into account. We therefore present a new R package for of data from the Illumina Infinium Human Methylation450 BeadChip (Illumina 450 K) built on concepts in recently published funNorm method, and introducing cell-type tissue-type flexibility. Results: funtooNorm is relevant sets containing samples two...
Abstract The recognition that individual GPCRs can activate multiple signaling pathways has raised the possibility of developing drugs selectively targeting therapeutically relevant ones. This requires tools to determine which G proteins and βarrestins are activated by a given receptor. Here, we present set BRET sensors monitoring activation 12 protein subtypes based on translocation their effectors plasma membrane (EMTA). Unlike most existing detection systems, EMTA does not require...
The study of complex heterodimeric peptide ligands has been hampered by a paucity pharmacological tools. To facilitate such investigations, we have explored the utility membrane tethered (MTLs). Feasibility this recombinant approach was with focus on <i>Drosophila</i> bursicon, cystine-knot protein that activates G protein–coupled receptor rickets (rk). Rk/bursicon signaling is an evolutionarily conserved pathway in insects required for wing expansion, cuticle hardening, and melanization...
Abstract Urocortin 2 (UCN2) acts as a ligand for the G protein-coupled receptor corticotropin-releasing hormone (CRHR2). UCN2 has been reported to improve or worsen insulin sensitivity and glucose tolerance in vivo. Here we show that acute dosing of induces systemic resistance male mice skeletal muscle. Inversely, chronic elevation by injection with adenovirus encoding resolves metabolic complications, improving tolerance. CRHR2 recruits Gs response low concentrations UCN2, well Gi...
Abstract Genome-wide loss-of-function screens using the CRISPR/Cas9 system allow efficient discovery of cancer cell vulnerabilities. While several studies have focused on correcting for DNA cleavage toxicity biases associated with copy number alterations, effects sgRNAs co-targeting multiple genomic loci in CRISPR not been discussed yet. In this work, we analyze essentiality screen data from 391 lines to characterize induced by multi-target sgRNAs. We investigate two types multi-targets:...
ABSTRACT Macrophages adopt dynamic cell states with distinct effector functions to maintain tissue homeostasis and respond environmental challenges. During chronic inflammation, macrophage polarization is subverted towards sustained inflammatory which contribute disease, but there limited understanding of the regulatory mechanisms underlying these disease-associated states. Here, we describe a systematic functional genomics approach that combines genome-wide phenotypic screening in primary...
Abstract Cas12a is a gene-editing tool that simplifies multiplexed gene targeting through its RNase activity, enabling maturation of individual crRNAs from pre-crRNA-encoding RNA. Here, we present mouse model constitutively expresses enhanced Acidaminococcus sp. ( enAsCas12a ) linked to an mCherry fluorescent reporter. We demonstrate efficient single and in cells KI mice. To test vivo transduced haematopoietic stem E μ -Myc T/+ ;enAsCas12a KI/+ animals with Trp53 -targeting pre-crRNAs...
<h3>Background</h3> A large fraction of patients with cancer exhibit an intrinsic or acquired resistance to immunotherapies that aim at boosting T cell responses against tumor cells. <h3>Methods</h3> To better understand mechanisms in cells, we used CRISPR-Cas9 whole-genome editing identify modified cells are resistant killing. <h3>Results</h3> Among the top hits, identified transcription factor zinc-finger protein X-linked (Zfx). Zfx knockout killing both <i>in vitro</i> and vivo</i>....
Anti-sense oligonucleotides (ASOs) are modified synthetic single-stranded molecules with enhanced stability, activity, and bioavailability. They associate RNA through sequence complementarity can reduce or alter mRNA expression upon binding of splice site positions. To target in the nucleus cytoplasm, ASOs must cross membranes, a poorly understood process. We have performed an unbiased CRISPR/Cas9 knockout screen genetic reporter to identify genes that increase decrease resulting most...
Abstract The success of CRISPR-mediated gene perturbation studies is highly dependent on the quality gRNAs, and several tools have been developed to enable optimal gRNA design. However, these are not all adaptable latest CRISPR modalities or nucleases, nor do they offer comprehensive annotation methods for advanced applications. Here, we present a new ecosystem R packages, called crispr-Verse , that enables efficient design multitude technologies. This includes knockout (CRISPRko),...