An entropy based heuristic model for predicting functional sub-type divisions of protein families

Multiple sequence alignments of protein families are often used for locating residues that are widely apart in the sequence, which are considered as influential for determining functional specificity of proteins towards various substrates, ligands, DNA and other proteins. In this paper, we propose an entropy-score based heuristic algorithm model for predicting functional sub-family divisions of protein families, given the multiple sequence alignment of the protein family as input without any functional sub-type or key site information given for any protein sequence. Two of the experimented test-cases are reported in this paper. First test-case is Nucleotidyl Cyclase protein family consisting of guanalyate and adenylate cyclases. And the second test-case is a dataset of proteins taken from six superfamilies in Structure-Function Linkage Database (SFLD). Results from these test-cases are reported in terms of confirmed sub-type divisions with phylogeny relations from former studies in the literature

Traversing the k-mer Landscape of NGS Read Datasets for Quality Score Sparsification

It is becoming increasingly impractical to indefinitely store raw sequencing data for later processing in an uncompressed state. In this paper, we describe a scalable compressive framework, Read-Quality-Sparsifier (RQS), which substantially outperforms the compression ratio and speed of other de novo quality score compression methods while maintaining SNP-calling accuracy. Surprisingly, RQS also improves the SNP-calling accuracy on a gold-standard, real-life sequencing dataset (NA12878) using a k-mer density profile constructed from 77 other individuals from the 1000 Genomes Project. This improvement in downstream accuracy emerges from the observation that quality score values within NGS datasets are inherently encoded in the k-mer landscape of the genomic sequences. To our knowledge, RQS is the first scalable sequence-based quality compression method that can efficiently compress quality scores of terabyte-sized and larger sequencing datasets. Availability: An implementation of our method, RQS, is available for download at: http://rqs.csail.mit.edu/. © 2014 Springer International Publishing Switzerland. Keywords: RQS; quality score; sparsification; compression; accuracy; variant callingHertz FoundationNational Institutes of Health (U.S.) (R01GM108348

HapTree-X: An Integrative Bayesian Framework for Haplotype Reconstruction from Transcriptome and Genome Sequencing Data

By running standard genotype calling tools, it is possible to accurately identify the number of wild type and mutant alleles for each single-nucleotide polymorphism (SNP) site. However, in the case of two heterozygous SNP sites, genotype calling tools cannot determine whether mutant alleles from different SNP loci are on the same chromosome or on different homologous chromosomes (i.e. compound heterozygote)

Fast genotyping of known SNPs through approximate

Motivation: As the volume of next-generation sequencing (NGS) data increases, faster algorithms become necessary. Although speeding up individual components of a sequence analysis pipeline (e.g. read mapping) can reduce the computational cost of analysis, such approaches do not take full advantage of the particulars of a given problem. One problem of great interest, genotyping a known set of variants (e.g. dbSNP or Affymetrix SNPs), is important for characterization of known genetic traits and causative disease variants within an individual, as well as the initial stage of many ancestral and population genomic pipelines (e.g. GWAS). Results: We introduce lightweight assignment of variant alleles (LAVA), an NGS-based genotyping algorithm for a given set of SNP loci, which takes advantage of the fact that approximate matching of mid-size k-mers (with k = 32) can typically uniquely ide ntify loci in the human genome without full read alignment. LAVA accurately calls the vast majority of SNPs in dbSNP and Affymetrix's Genome-Wide Human SNP Array 6.0 up to about an order of magnitude faster than standard NGS genotyping pipelines. For Affymetrix SNPs, LAVA has significantly higher SNP calling accuracy than existing pipelines while using as low as ∼5 GB of RAM. As such, LAVA represents a scalable computational method for population-level genotyping studies as well as a flexible NGS-based replacement for SNP arrays. Availability and Implementation: LAVA software is available at http://lava.csail.mit.edu

Next-generation VariationHunter: combinatorial algorithms for transposon insertion discovery

Recent years have witnessed an increase in research activity for the detection of structural variants (SVs) and their association to human disease. The advent of next-generation sequencing technologies make it possible to extend the scope of structural variation studies to a point previously unimaginable as exemplified by the 1000 Genomes Project. Although various computational methods have been described for the detection of SVs, no such algorithm is yet fully capable of discovering transposon insertions, a very important class of SVs to the study of human evolution and disease. In this article, we provide a complete and novel formulation to discover both loci and classes of transposons inserted into genomes sequenced with high-throughput sequencing technologies. In addition, we also present ‘conflict resolution’ improvements to our earlier combinatorial SV detection algorithm (VariationHunter) by taking the diploid nature of the human genome into consideration. We test our algorithms with simulated data from the Venter genome (HuRef) and are able to discover >85% of transposon insertion events with precision of >90%. We also demonstrate that our conflict resolution algorithm (denoted as VariationHunter-CR) outperforms current state of the art (such as original VariationHunter, BreakDancer and MoDIL) algorithms when tested on the genome of the Yoruba African individual (NA18507)

HapTree: A Novel Bayesian Framework for Single Individual Polyplotyping Using NGS Data

As the more recent next-generation sequencing (NGS) technologies provide longer read sequences, the use of sequencing datasets for complete haplotype phasing is fast becoming a reality, allowing haplotype reconstruction of a single sequenced genome. Nearly all previous haplotype reconstruction studies have focused on diploid genomes and are rarely scalable to genomes with higher ploidy. Yet computational investigations into polyploid genomes carry great importance, impacting plant, yeast and fish genomics, as well as the studies of the evolution of modern-day eukaryotes and (epi)genetic interactions between copies of genes. In this paper, we describe a novel maximum-likelihood estimation framework, HapTree, for polyploid haplotype assembly of an individual genome using NGS read datasets. We evaluate the performance of HapTree on simulated polyploid sequencing read data modeled after Illumina sequencing technologies. For triploid and higher ploidy genomes, we demonstrate that HapTree substantially improves haplotype assembly accuracy and efficiency over the state-of-the-art; moreover, HapTree is the first scalable polyplotyping method for higher ploidy. As a proof of concept, we also test our method on real sequencing data from NA12878 (1000 Genomes Project) and evaluate the quality of assembled haplotypes with respect to trio-based diplotype annotation as the ground truth. The results indicate that HapTree significantly improves the switch accuracy within phased haplotype blocks as compared to existing haplotype assembly methods, while producing comparable minimum error correction (MEC) values. A summary of this paper appears in the proceedings of the RECOMB 2014 conference, April 2–5.National Science Foundation (U.S.) (NSF/NIH BIGDATA Grant R01GM108348-01)National Science Foundation (U.S.) (Graduate Research Fellowship)Simons Foundatio

Scalable methods for storage, processing and analysis of sequencing datasets

Thesis: Ph. D., Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science, 2017.Cataloged from PDF version of thesis.Includes bibliographical references (pages 179-189).Massive amounts of next-generation sequencing (NGS) reads generated from sequencing machines around the world have revolutionized biotechnology enabling wide-scale disease and variation studies, personalized medicine and helping us understand our evolutionary history. However, the amount of sequencing data generated every day increases at an exponential rate posing an imminent need for smart algorithmic solutions to handle massive sequencing datasets and efficiently extract the useful knowledge within them. This thesis consists of four research contributions on these two fronts. First, we present a computational framework that leverages the redundancy within large genomic datasets for performing faster read-mapping while improving sensitivity. Second, we describe a lossy compression method for quality scores within sequencing datasets that strikingly improves the downstream accuracy for genotyping. Third, we introduce a Bayesian framework for accurate diploid and polyploid haplotype reconstruction of an individual genome using NGS datasets. Lastly, we extend this haplotype reconstruction framework to high-throughput transcriptome sequencing datasets.by Deniz Yorukoglu.Ph. D

Detection and characterization of novel structural alterations in transcribed sequences

One of the key problems in computational genomics is that of identifying structural variations between two sequences of genomic origin. Recently, with the advent of high-throughput sequencing of transcriptomes (RNA-seq), transcriptional structural variation studies also came into prominence. This study introduces two novel frameworks for aligning transcribed sequences to the genome with high sensitivity to structural alterations within the transcript. (1) A pairwise nucleotide-level alignment model and (2) a faster lower-sensitivity solution based on chaining homologous substrings between the transcript and the genome. A further contribution of this study is a stand-alone transcriptome-to-genome alignment tool, which can comprehensively identify and characterize transcriptional events (duplications, inversions, rearrangements and fusions); suitable for high-throughput structural variation studies involving long transcribed sequences with high similarity to their genomic origin. Reported results include experiments upon simulated datasets of transcriptional events and RNA-seq assemblies of a human prostate cancer individual