5,600 research outputs found
Modeling ChIP Sequencing In Silico with Applications
ChIP sequencing (ChIP-seq) is a new method for genomewide mapping of protein binding sites on DNA. It has generated much excitement in functional genomics. To score data and determine adequate sequencing depth, both the genomic background and the binding sites must be properly modeled. To develop a computational foundation to tackle these issues, we first performed a study to characterize the observed statistical nature of this new type of high-throughput data. By linking sequence tags into clusters, we show that there are two components to the distribution of tag counts observed in a number of recent experiments: an initial power-law distribution and a subsequent long right tail. Then we develop in silico ChIP-seq, a computational method to simulate the experimental outcome by placing tags onto the genome according to particular assumed distributions for the actual binding sites and for the background genomic sequence. In contrast to current assumptions, our results show that both the background and the binding sites need to have a markedly nonuniform distribution in order to correctly model the observed ChIP-seq data, with, for instance, the background tag counts modeled by a gamma distribution. On the basis of these results, we extend an existing scoring approach by using a more realistic genomic-background model. This enables us to identify transcription-factor binding sites in ChIP-seq data in a statistically rigorous fashion
Detection of regulator genes and eQTLs in gene networks
Genetic differences between individuals associated to quantitative phenotypic
traits, including disease states, are usually found in non-coding genomic
regions. These genetic variants are often also associated to differences in
expression levels of nearby genes (they are "expression quantitative trait
loci" or eQTLs for short) and presumably play a gene regulatory role, affecting
the status of molecular networks of interacting genes, proteins and
metabolites. Computational systems biology approaches to reconstruct causal
gene networks from large-scale omics data have therefore become essential to
understand the structure of networks controlled by eQTLs together with other
regulatory genes, and to generate detailed hypotheses about the molecular
mechanisms that lead from genotype to phenotype. Here we review the main
analytical methods and softwares to identify eQTLs and their associated genes,
to reconstruct co-expression networks and modules, to reconstruct causal
Bayesian gene and module networks, and to validate predicted networks in
silico.Comment: minor revision with typos corrected; review article; 24 pages, 2
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Patient and Disease-Specific Induced Pluripotent Stem Cells for Discovery of Personalized Cardiovascular Drugs and Therapeutics.
Human induced pluripotent stem cells (iPSCs) have emerged as an effective platform for regenerative therapy, disease modeling, and drug discovery. iPSCs allow for the production of limitless supply of patient-specific somatic cells that enable advancement in cardiovascular precision medicine. Over the past decade, researchers have developed protocols to differentiate iPSCs to multiple cardiovascular lineages, as well as to enhance the maturity and functionality of these cells. Despite significant advances, drug therapy and discovery for cardiovascular disease have lagged behind other fields such as oncology. We speculate that this paucity of drug discovery is due to a previous lack of efficient, reproducible, and translational model systems. Notably, existing drug discovery and testing platforms rely on animal studies and clinical trials, but investigations in animal models have inherent limitations due to interspecies differences. Moreover, clinical trials are inherently flawed by assuming that all individuals with a disease will respond identically to a therapy, ignoring the genetic and epigenomic variations that define our individuality. With ever-improving differentiation and phenotyping methods, patient-specific iPSC-derived cardiovascular cells allow unprecedented opportunities to discover new drug targets and screen compounds for cardiovascular disease. Imbued with the genetic information of an individual, iPSCs will vastly improve our ability to test drugs efficiently, as well as tailor and titrate drug therapy for each patient
Basic and applied uses of genome-scale metabolic network reconstructions of Escherichia coli.
The genome-scale model (GEM) of metabolism in the bacterium Escherichia coli K-12 has been in development for over a decade and is now in wide use. GEM-enabled studies of E. coli have been primarily focused on six applications: (1) metabolic engineering, (2) model-driven discovery, (3) prediction of cellular phenotypes, (4) analysis of biological network properties, (5) studies of evolutionary processes, and (6) models of interspecies interactions. In this review, we provide an overview of these applications along with a critical assessment of their successes and limitations, and a perspective on likely future developments in the field. Taken together, the studies performed over the past decade have established a genome-scale mechanistic understanding of genotype–phenotype relationships in E. coli metabolism that forms the basis for similar efforts for other microbial species. Future challenges include the expansion of GEMs by integrating additional cellular processes beyond metabolism, the identification of key constraints based on emerging data types, and the development of computational methods able to handle such large-scale network models with sufficient accuracy
Inference of Markovian Properties of Molecular Sequences from NGS Data and Applications to Comparative Genomics
Next Generation Sequencing (NGS) technologies generate large amounts of short
read data for many different organisms. The fact that NGS reads are generally
short makes it challenging to assemble the reads and reconstruct the original
genome sequence. For clustering genomes using such NGS data, word-count based
alignment-free sequence comparison is a promising approach, but for this
approach, the underlying expected word counts are essential.
A plausible model for this underlying distribution of word counts is given
through modelling the DNA sequence as a Markov chain (MC). For single long
sequences, efficient statistics are available to estimate the order of MCs and
the transition probability matrix for the sequences. As NGS data do not provide
a single long sequence, inference methods on Markovian properties of sequences
based on single long sequences cannot be directly used for NGS short read data.
Here we derive a normal approximation for such word counts. We also show that
the traditional Chi-square statistic has an approximate gamma distribution,
using the Lander-Waterman model for physical mapping. We propose several
methods to estimate the order of the MC based on NGS reads and evaluate them
using simulations. We illustrate the applications of our results by clustering
genomic sequences of several vertebrate and tree species based on NGS reads
using alignment-free sequence dissimilarity measures. We find that the
estimated order of the MC has a considerable effect on the clustering results,
and that the clustering results that use a MC of the estimated order give a
plausible clustering of the species.Comment: accepted by RECOMB-SEQ 201
Methodological Issues in Multistage Genome-Wide Association Studies
Because of the high cost of commercial genotyping chip technologies, many
investigations have used a two-stage design for genome-wide association
studies, using part of the sample for an initial discovery of ``promising''
SNPs at a less stringent significance level and the remainder in a joint
analysis of just these SNPs using custom genotyping. Typical cost savings of
about 50% are possible with this design to obtain comparable levels of overall
type I error and power by using about half the sample for stage I and carrying
about 0.1% of SNPs forward to the second stage, the optimal design depending
primarily upon the ratio of costs per genotype for stages I and II. However,
with the rapidly declining costs of the commercial panels, the generally low
observed ORs of current studies, and many studies aiming to test multiple
hypotheses and multiple endpoints, many investigators are abandoning the
two-stage design in favor of simply genotyping all available subjects using a
standard high-density panel. Concern is sometimes raised about the absence of a
``replication'' panel in this approach, as required by some high-profile
journals, but it must be appreciated that the two-stage design is not a
discovery/replication design but simply a more efficient design for discovery
using a joint analysis of the data from both stages. Once a subset of
highly-significant associations has been discovered, a truly independent
``exact replication'' study is needed in a similar population of the same
promising SNPs using similar methods.Comment: Published in at http://dx.doi.org/10.1214/09-STS288 the Statistical
Science (http://www.imstat.org/sts/) by the Institute of Mathematical
Statistics (http://www.imstat.org
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