A5040631498- Genomics and Chromatin Dynamics
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Chromosomal and Genetic Variations
- Genetics, Aging, and Longevity in Model Organisms
- Animal Genetics and Reproduction
- Developmental Biology and Gene Regulation
- Epigenetics and DNA Methylation
- Plant Molecular Biology Research
- Reproductive Biology and Fertility
- Insect symbiosis and bacterial influences
- Pluripotent Stem Cells Research
- Academic Publishing and Open Access
- DNA Repair Mechanisms
- Genetic and Clinical Aspects of Sex Determination and Chromosomal Abnormalities
- Insect Resistance and Genetics
- RNA Interference and Gene Delivery
- Protein Degradation and Inhibitors
- Gene Regulatory Network Analysis
- Chromatin Remodeling and Cancer
- Microtubule and mitosis dynamics
- Research Data Management Practices
- Light effects on plants
- Poverty, Education, and Child Welfare
- Invertebrate Immune Response Mechanisms
University of Wisconsin–Madison
2015-2024
European Bioinformatics Institute
2024
eLife Sciences Publications
2017
Rochester Institute of Technology
2016
Madison Group (United States)
2013
University of California, Berkeley
2010-2011
Howard Hughes Medical Institute
2005-2011
Massachusetts Institute of Technology
2005-2011
Florida Fish and Wildlife Conservation Commission
2005
Kansas State University
1993
Abstract We have adapted a bacterial CRISPR RNA/Cas9 system to precisely engineer the Drosophila genome and report that Cas9-mediated genomic modifications are efficiently transmitted through germline. This RNA-guided Cas9 can be rapidly programmed generate targeted alleles for probing gene function in Drosophila.
The earliest stages of development in most metazoans are driven by maternally deposited proteins and mRNAs, with widespread transcriptional activation the zygotic genome occurring hours after fertilization, at a period known as maternal-to-zygotic transition (MZT). In Drosophila, MZT is preceded transcription small number genes that initiate sex determination, patterning, other early developmental processes; zinc-finger protein Zelda (ZLD) plays key role their activation. To better...
Multiple stringent confinement strategies should be used whenever possible
We describe the genome-wide distributions and temporal dynamics of nucleosomes post-translational histone modifications throughout maternal-to-zygotic transition in embryos Drosophila melanogaster. At mitotic cycle 8, when few zygotic genes are being transcribed, embryonic chromatin is a relatively simple state: there nucleosome free regions, undetectable levels methylation marks characteristic mature chromatin, low acetylation at small number loci. Histone increases by 12, but it not until...
Abstract The CRISPR‐Cas9 system has transformed genome engineering of model organisms from possible to practical. can be readily programmed generate sequence‐specific double‐strand breaks that disrupt targeted loci when repaired by error‐prone non‐homologous end joining (NHEJ) or catalyze precise modification through homology‐directed repair (HDR). Here we describe a streamlined approach for rapid and highly efficient the Drosophila via CRISPR‐Cas9‐mediated HDR. In this approach, transgenic...
The regulation of transcription requires the coordination numerous activities on DNA, yet how factors mediate these remains poorly understood. Here, we use lattice light-sheet microscopy to integrate single-molecule and high-speed 4D imaging in developing Drosophila embryos study nuclear organization interactions key Zelda Bicoid. In contrast previous studies suggesting stable, cooperative binding, show that both interact with DNA surprisingly high off-rates. We find form dynamic subnuclear...
The transition from a specified germ cell to population of pluripotent cells occurs rapidly following fertilization. During this developmental transition, the zygotic genome is largely transcriptionally quiescent and undergoes significant chromatin remodeling. In Drosophila , DNA-binding protein Zelda (also known as Vielfaltig) required for transcriptional activation genome. Open associated with Zelda-bound loci, well more generally regions active transcription. Nonetheless, extent which...
Following fertilization, the genomes of germ cells are reprogrammed to form totipotent embryo. Pioneer transcription factors essential for remodeling chromatin and driving initial wave zygotic gene expression. In Drosophila melanogaster , pioneer factor Zelda is development through this dramatic period reprogramming, known as maternal-to-zygotic transition (MZT). However, it was unknown whether additional were required transition. We identified an maternally encoded MZT, GAGA Factor (GAF)....
Abstract Pioneer transcription factors can engage nucleosomal DNA, which leads to local chromatin remodeling and the establishment of transcriptional competence. However, impact enhancer priming by pioneer on temporal control gene expression mitotic memory remains unclear. Here we employ quantitative live imaging methods mathematical modeling test effect factor Zelda dynamics in Drosophila embryos. We demonstrate that increasing number binding sites accelerates kinetics nuclei activation...
The CRISPR/Cas9 system has attracted significant attention for its potential to transform genome engineering. We and others have recently shown that the RNA-guided Cas9 nuclease can be employed engineer Drosophila genome, these modifications are efficiently transmitted through germline. A single targeting RNA guide a specific genomic sequence where it induces double-strand breaks that, when imperfectly repaired, yield mutations. also demonstrated 2 RNAs used generate large defined deletions...
Abstract To maintain cellular identities during development, gene expression profiles must be faithfully propagated through cell generations. The reestablishment of patterns upon mitotic exit is mediated, in part, by transcription factors (TF) bookmarking. However, the mechanisms and functions TF bookmarking early embryogenesis remain poorly understood. In this study, taking advantage naturally synchronized mitoses Drosophila embryos, we provide evidence that GAGA pioneer factor (GAF) acts...
The Caenorhabditis elegans synthetic multivulva (synMuv) genes act redundantly to antagonize the specification of vulval cell fates, which are promoted by an RTK/Ras pathway. At least 26 synMuv have been genetically identified, several encode proteins with homologs that in chromatin remodeling or transcriptional repression. Here we report molecular characterization two genes, lin-37 and lin-54. We show lin-54 a complex at seven proteins, including LIN-35, only C. homolog mammalian tumor...