Hybridation génomique comparative

L'hybridation génomique comparative (en anglais, Comparative Genomic Hybridization ou CGH) est une technique de cytogénétique moléculaire permettant d'analyser les variations du nombre de copies dans l'ADN. Principe Dans un organisme diploïde tel que l'humain, chaque segment d'ADN peut être trouvé en duplicat : une copie est présente sur chacun des deux chromosomes d'une paire. Dans certaines pathologies, le nombre de copies peut varier : il peut augmenter par exemple en cas de duplication et diminuer dans le cas de délétion. L'hybridation génomique comparative permet de diagnostiquer ces pathologies et d'identifier les régions chromosomiques impliquées. Méthode ; Marquage des sondes L'ADN d'un patient est extrait de tissu normal (souvent des lymphocytes) et de tissu potentiellement tumoral puis marqué par des fluorochromes. En général, l'ADN tumoral est marqué en vert par la fluorescéine (sous forme de dUTP-FITC) et l'ADN de tissu sain en rouge par la rhod

The Gene Family-Free Median of Three

Metin Balaban

The gene family-free framework for comparative genomics aims at developing methods for gene order analysis that do not require prior gene family assignment, but work directly on a sequence similarity graph. We present a model for constructing a median of three genomes in this family-free setting, based on maximizing an objective function that generalizes the classical breakpoint distance by integrating sequence similarity in the score of a gene adjacency. We show that the corresponding computational problem is MAX SNP-hard and we present a 0-1 linear program for its exact solution. The result of this program is a median genome with median genes associated to extant genes, in which median adjacencies are assumed to define positional orthologs. We demonstrate through simulations and comparison with the OMA orthology database that the herein presented method is able compute accurate medians and positional orthologs for genomes comparable in size of bacterial genomes.

Springer AG2016

Improving Comparative Genomic Studies

Cristina Gabriela Ghiurcuta

Comparative genomics aims to understand the structure of genomes and the function of various genomic fragments, by transferring knowledge gained from well studied genomes, to the new object of study. Rapid and inexpensive high-throughput sequencing is making available more and more complete genome sequences. Despite the significant scientific advance, we still lack good models for the evolution of the genomic architecture, therefore analyzing these genomes presents formidable challenges. Early approaches used pairwise comparisons, but today researchers are attempting to leverage the larger potential of multiway comparisons. Current approaches are based on the identification of so called syntenic blocks: blocks of sequence that exhibit conserved features across the genomes under study. Syntenic blocks are in many ways analogous to genesâ -in many cases, the markers are used to constructing them are genes. Like genes they can exist in multiple copies, in which case we could define analogs of orthology and paralogy. However, whereas genes are studied at the sequence level, syntenic blocks are too large for that level of detail - it is their structure and function as a unit that makes them valuable for genome level comparative studies. Syntenic blocks are required for complex computations to scale to the billions of nucleotides present in many genomes; they enable comparisons across broad ranges of genomes because they filter outmuch of the individual variability; they highlight candidate regions for in-depth studies; and they facilitate whole-genome comparisons through visualization tools. The identification of such blocks is the first step in comparative studies, yet its effect on final results has not been well studied, nor has any formalization of syntenic blocks been proposed. Tools for the identification of syntenic blocks yield quite different results, thereby preventing a systematic assessment of the next steps in an analysis. Current tools do not include measurable quality objectives and thus cannot be benchmarked against themselves. Comparisons among tools have also been neglected - what few results are given use superficial measures unrelated to quality or consistency. In this thesis we address two major challenges, and present: (i) a theoretical model as well as an experimental basis for comparing syntenic blocks and thus also for improving the design of tools for the identification of syntenic blocks; (ii) a prototype model that serves as a basis for implementing effective synteny mining tools. We offer an overview of the milestones present in literature, on the development of concepts and tool related to synteny; we illustrate the application of the model and the measures by applying them to syntenic blocks produced by different contemporary tools on publicly available data sets. We have taken the first step towards a formal approach to the construction of syntenic blocks by developing a simple quality criterion based on sound evolutionary principles. Our experiments demonstrate widely divergent results among these tools, throwing into question the robustness of the basic approach in comparative genomics. Our findings highlight the need for a well founded, systematic approach to the decomposition of genomes into syntenic blocks and motivate the second part of the work - starting from the proposed model, we extend the concept with data dependent features and constraints, in order to test the concept on cases of interest.

EPFL2014

Improving Comparative Genomic Studies

Cristina Gabriela Ghiurcuta

EPFL2014