Recent Publications

Development. 2012 Oct;139(19):3613-22. doi: 10.1242/dev.081828.

ISWI contributes to ArsI insulator function in development of the sea urchin.
Yajima M, Fairbrother WG, Wessel GM.
Source
MCB Department, Brown University, 185 Meeting Street, BOX-GL173, Providence, RI 02912, USA. mamiko_yajima@brown.edu
Abstract
Insulators are genomic elements that regulate transcriptional activity by forming chromatin boundaries. Various DNA insulators have been identified or are postulated in many organisms, and the paradigmatic CTCF-dependent insulators are perhaps the best understood and most widespread in function. The diversity of DNA insulators is, however, understudied, especially in the context of embryonic development, when many new gene territories undergo transitions in functionality. Here we report the functional analysis of the arylsulfatase insulator (ArsI) derived from the sea urchin, which has conserved insulator activity throughout the many metazoans tested, but for which the molecular mechanism of function is unknown. Using a rapid in vivo assay system and a high-throughput mega-shift assay, we identified a minimal region in ArsI that is responsible for its insulator function. We discovered a small set of proteins specifically bound to the minimal ArsI region, including ISWI, a known chromatin-remodeling protein. During embryogenesis, ISWI was found to interact with select ArsI sites throughout the genome, and when inactivated led to misregulation of select gene expression, loss of insulator activity and aberrant morphogenesis. These studies reveal a mechanistic basis for ArsI function in the gene regulatory network of early development.
PMID:
 
22949616
 
[PubMed - indexed for MEDLINE] 
PMCID:
 
PMC3436113
 [Available on 2013/10/1

NATURE STRUCTURAL & MOLECULAR BIOLOGY | BRIEF COMMUNICATION

Large-scale mapping of branchpoints in human pre-mRNA transcripts in vivo



Nature Structural & Molecular Biology
 
(2012)
 
doi:10.1038/nsmb.2327
Received
 
23 February 2012 
Accepted
 
17 May 2012 
Published online
 
17 June 2012

We present the first large-scale identification of lariats—the transient branched introns that are released as a byproduct of pre-mRNA splicing. The locations of the branchpoints in these introns provide insight into the early steps of splicing. From this data set, we have developed a comprehensive model of 3′ splice-site selection, identified new mechanisms of alternative splicing and mapped the distribution of splicing factors around branchpoints.



Genomics. 2011 Oct 1. [Epub ahead of print]

Genome-wide transcriptome analysis in murine neural retina using high-throughput RNA sequencing.

Abstract

Genome-wide characterization of the retinal transcriptome is central to understanding development, physiology and disorders of the visual system. Massively parallel, short-read sequencing of mRNA libraries was used to generate an extensive map of the transcriptome of the adult, murine neural retina. RNA-seq data strongly corroborates prior transcriptome studies by microarray and SAGE. However, several novel features of the retinal transcriptome were discovered. For example, retinal disease genes were discovered to be among the most highly expressed in the transcriptome. We also demonstrate other interesting features of the retinal transcriptome, for example, that the retina appears to employ a very specific and restricted set of synaptic vesicle genes, and also that there is persistence of expression of a majority of "neurodevelopmental" genes into adulthood. Retina transcriptome studies utilizing novel sequencing methods have been highly informative and these data may also serve as a resource for the community of researchers.

Copyright © 2011. Published by Elsevier Inc.

PMID:
22032952
[PubMed - as supplied by publisher]

J Biol Chem. 2011 Sep 27. [Epub ahead of print]

The transcriptome of a human polar body accurately reflects its sibling oocyte.

Source

Brown University, United States;

Abstract


Improved methods are needed to reliably and accurately evaluate oocyte quality prior to fertilization and transfer into the woman of human embryos created through in vitro fertilization (IVF). All oocytes that are retrieved and mature in culture are exposed to sperm with little in the way of evaluating the oocyte quality. Further, embryos created through IVF are currently evaluated for developmental potential by morphology, a criterion lacking in quantitation and accuracy. With the recent successes in oocyte vitrification and storage, clear metrics are needed to determine oocyte quality prior to fertilizing.

PMID:
 
21953461
 
[PubMed - as supplied by publisher]

Differential Gene Expression in the Siphonophore Nanomia bijuga (Cnidaria) Assessed with Multiple Next-Generation Sequencing Workflows


Stefan Siebert1*, Mark D. Robinson2,3, Sophia C. Tintori1, Freya Goetz1, Rebecca R. Helm1, Stephen A. Smith1,4, Nathan Shaner5, Steven H. D. Haddock5, Casey W. Dunn1*

1 Department of Ecology and Evolutionary Biology, Brown University, Providence, Rhode Island, United States of America, 2 Epigenetics Laboratory, Cancer Research Program, Garvan Institute of Medical Research, Sydney, New South Wales, Australia, 3 Bioinformatics Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia, 4 Heidelberg Institute for Theoretical Studies, Heidelberg, Germany, 5 Monterey Bay Aquarium Research Institute, Moss Landing, California, United States of America

Abstract 

We investigated differential gene expression between functionally specialized feeding polyps and swimming medusae in the siphonophore Nanomia bijuga (Cnidaria) with a hybrid long-read/short-read sequencing strategy. We assembled a set of partial gene reference sequences from long-read data (Roche 454), and generated short-read sequences from replicated tissue samples that were mapped to the references to quantify expression. We collected and compared expression data with three short-read expression workflows that differ in sample preparation, sequencing technology, and mapping tools. These workflows were Illumina mRNA-Seq, which generates sequence reads from random locations along each transcript, and two tag-based approaches, SOLiD SAGE and Helicos DGE, which generate reads from particular tag sites. Differences in expression results across workflows were mostly due to the differential impact of missing data in the partial reference sequences. When all 454-derived gene reference sequences were considered, Illumina mRNA-Seq detected more than twice as many differentially expressed (DE) reference sequences as the tag-based workflows. This discrepancy was largely due to missing tag sites in the partial reference that led to false negatives in the tag-based workflows. When only the subset of reference sequences that unambiguously have tag sites was considered, we found broad congruence across workflows, and they all identified a similar set of DE sequences. Our results are promising in several regards for gene expression studies in non-model organisms. First, we demonstrate that a hybrid long-read/short-read sequencing strategy is an effective way to collect gene expression data when an annotated genome sequence is not available. Second, our replicated sampling indicates that expression profiles are highly consistent across field-collected animals in this case. Third, the impacts of partial reference sequences on the ability to detect DE can be mitigated through workflow choice and deeper reference sequencing.

Received: April 6, 2011; Accepted: July 1, 2011; Published: July 29, 2011


Resolving the evolutionary relationships of molluscs with phylogenomic tools


Nature
 
(2011)
 
doi:10.1038/nature10526
Received
 
07 April 2011
 
Accepted
 
31 August 2011
 
Published online
 
26 October 2011

Molluscs (snails, octopuses, clams and their relatives) have a great disparity of body plans and, among the animals, only arthropods surpass them in species number. This diversity has made Mollusca one of the best-studied groups of animals, yet their evolutionary relationships remain poorly resolved1. Open questions have important implications for the origin of Mollusca and for morphological evolution within the group. These questions include whether the shell-less, vermiform aplacophoran molluscs diverged before the origin of the shelled molluscs (Conchifera)234 or lost their shells secondarily. Monoplacophorans were not included in molecular studies until recently56, when it was proposed that they constitute a clade named Serialia together with Polyplacophora (chitons), reflecting the serial repetition of body organs in both groups5. Attempts to understand the early evolution of molluscs become even more complex when considering the large diversity of Cambrian fossils. These can have multiple dorsal shell plates and sclerites78910 or can be shell-less but with a typical molluscan radula and serially repeated gills11. To better resolve the relationships among molluscs, we generated transcriptome data for 15 species that, in combination with existing data, represent for the first time all major molluscan groups. We analysed multiple data sets containing up to 216,402 sites and 1,185 gene regions using multiple models and methods. Our results support the clade Aculifera, containing the three molluscan groups with spicules but without true shells, and they support the monophyly of Conchifera. Monoplacophora is not the sister group to other Conchifera but to Cephalopoda. Strong support is found for a clade that comprises Scaphopoda (tusk shells), Gastropoda and Bivalvia, with most analyses placing Scaphopoda and Gastropoda as sister groups. This well-resolved tree will constitute a framework for further studies of mollusc evolution, development and anatomy.


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