A5101563397- Microtubule and mitosis dynamics
- Genomics and Chromatin Dynamics
- Chromosomal and Genetic Variations
- Cell Image Analysis Techniques
- Single-cell and spatial transcriptomics
- Nuclear Structure and Function
- CRISPR and Genetic Engineering
- Image Processing Techniques and Applications
- RNA Interference and Gene Delivery
- Lymphoma Diagnosis and Treatment
- Cellular Mechanics and Interactions
- Electron and X-Ray Spectroscopy Techniques
- Gastrointestinal Tumor Research and Treatment
- Cancer-related Molecular Pathways
- Cell death mechanisms and regulation
- Digital Imaging for Blood Diseases
- Neurofibromatosis and Schwannoma Cases
- Advanced biosensing and bioanalysis techniques
- Nuclear Physics and Applications
- Silk-based biomaterials and applications
- MicroRNA in disease regulation
- Epigenetics and DNA Methylation
- Virus-based gene therapy research
- Bioinformatics and Genomic Networks
- Photosynthetic Processes and Mechanisms
Cairn Biosciences (United States)
2020-2024
Howard Hughes Medical Institute
2014-2021
University of California, San Francisco
2014-2021
University of Edinburgh
2006-2015
Marine Biological Laboratory
2015
Wellcome Centre for Cell Biology
2006-2014
University of California, Berkeley
2014
Johns Hopkins Medicine
2008
Johns Hopkins University
2008
Centre for Genomic Regulation
2008
A longstanding question in centromere biology has been the organization of CENP-A–containing chromatin and its implications for kinetochore assembly. Here, we have combined genetic manipulations with deconvolution super-resolution fluorescence microscopy a detailed structural analysis chicken kinetochores. Using subdiffraction spatial resolution single molecule sensitivity to map protein localization unfolded by exposure low salt buffer, observed robust amounts H3K9me3, but only levels...
Repo-Man targets protein phosphatase 1 γ (PP1γ) to chromatin at anaphase onset and regulates chromosome structure during mitotic exit. Here, we show that a Repo-Man:PP1 complex forms in following dephosphorylation of Repo-Man. Upon activation, the localizes chromosomes causes histone H3 (Thr3, Ser10, Ser28). In anaphase, has both catalytic structural functions are mediated by two separate domains. A C-terminal domain bulk early anaphase. There, it PP1 for possibly other chromosomal...
When chromosomes are aligned and bioriented at metaphase, the elastic stretch of centromeric chromatin opposes pulling forces exerted on sister kinetochores by mitotic spindle. Here we show that condensin ATPase activity is an important regulator centromere stiffness function. Condensin depletion decreases 50% when applied to kinetochores. However, dispensable for normal level compaction (rest length) centromeres, which probably depends other factors control higher-order folding....
CRISPR (clustered regularly interspaced short palindromic repeats)-based gene inactivation provides a powerful means for linking genes to particular cellular phenotypes. CRISPR-based screening typically uses large genomic pools of single guide RNAs (sgRNAs). However, this approach is limited phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed microscopy-based approach, which name optical enrichment, select cells displaying CRISPR-induced phenotype...
Survivin is a key cellular protein thought to function in apoptotic regulation, mitotic progression, or possibly both. In this study, we describe the isolation of two conditional knockouts survivin gene chicken DT40 cells. cells lacking die interphase after failing complete cytokinesis. However, these show normal sensitivity chemotherapeutic agent etoposide. Expression mutants against null background reassess role several residues reveals that can grow normally if their sole missing widely...
The formation of the mitotic spindle is a complex process that requires massive cellular reorganization. Regulation by kinases controls this entire process. One these controllers Aurora A kinase, which itself highly regulated. In study, we show nuclear pore protein ALADIN novel spatial regulator A. Without ALADIN, spreads from centrosomes onto microtubules, affects distribution subset microtubule regulators and slows assembly chromosome alignment. interacts with inactive recruited to pole...
We describe a method for the isolation of conditional knockouts essential multiply spliced genes in which entire body gene downstream ATG start codon is left untouched but can be switched off rapidly and completely by adding tetracycline to culture medium. The approach centers on "promoter-hijack" strategy gene's promoter replaced with minimal responsive tetracycline-repressible transactivator (tTA). Elsewhere genome, cloned fragment used drive expression tTA. Thus, essentially regulated its...
Gliosarcomas are rare and poorly characterized malignant brain tumors that exhibit a biphasic tissue pattern with areas of gliomatous sarcomatous differentiation. These histological variants glioblastoma, displaying similar genetic profile dismal prognosis. Up‐regulation PDGFR subfamily tyrosine kinase members, PDGFR‐ α c‐Kit, their intracellular effectors RAS/RAF/MAPK has crucial role in the cancer development. In addition, signal transduction mediated by activating mutations c ‐Kit can be...
Large multinucleated Reed-Sternberg cells (RS) and large mononucleated Hodgkin (H) are traditionally considered to be the neoplastic population in classical lymphoma, (cHL) postulated promote disease. However, contribution of these larger progression cHL remains debatable. We used established cell lines cellular fractions composed small only or enriched RS/H investigate origin characterize which they derive from. confirm that give rise cells, we show latter proliferate significantly more...
Focal adhesions are dynamic structures that interact with the extracellular matrix on cell exterior and actin filaments interior, enabling cells to adhere crawl along surfaces. We describe a system for inducing formation of focal in normally non–ECM-adherent, nonmotile Drosophila S2 cells. These contain expected molecular markers such as talin, vinculin, p130Cas, they require talin their formation. The induced also display nonpolarized form motility vitronectin-coated substrates. Consistent...
In order to understand the three-dimensional structure of functional kinetochore in vertebrates, we require a complete list and stoichiometry for protein components kinetochore, which can be provided by genetic proteomic experiments. We also need know how chromatin-containing CENP-A, makes up structural foundation is folded, much that DNA involved assembling kinetochore. this MS, demonstrate functioning metaphase kinetochores chicken DT40 cells contain roughly 50 kb DNA, an amount...
ABSTRACT CRISPR (clustered regularly interspaced short palindromic repeats) -based gene inactivation provides a powerful means of linking genes to particular cellular phenotypes. CRISPR-based screening has mostly relied upon using large genomic pools single guide RNAs (sgRNAs). However, this approach is limited phenotypes that can be enriched by chemical selection or FACS sorting. Here, we developed microscopy-based approach, which name optical enrichment, computationally select cells...
Fluorescent live-cell microscopy is essential for understanding cellular dynamics by imaging specific molecules, their interactions, and biochemical states in live samples. It crucial biological research drug screening. However, often requires balancing temporal resolution with cell viability (i.e. conditions enabling cells to thrive) because of photo-toxicity. Consequently, there increasing interest lowering allow extended observation periods study event sequences that uncover causal...
Abstract High-content screening (HCS) provides an excellent tool to understand the mechanism of action drugs on disease-relevant model systems. Careful selection fluorescent labels (FLs) is crucial for successful HCS assay development. assays typically comprise (a) FLs containing biological information interest, and (b) additional structural enabling instance segmentation downstream analysis. However, limited number available fluorescence microscopy imaging channels restricts degree which...
Abstract High-content screening (HCS) provides an excellent tool to understand the mechanism of action drugs on disease-relevant model systems. Careful selection fluorescent labels (FLs) is crucial for successful HCS assay development. assays typically comprise (1) FLs containing biological information interest, and (2) additional structural enabling instance segmentation downstream analysis. However, limited number available fluorescence microscopy imaging channels restricts degree which...
Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – 7,