Prediction of regulatory motifs from human Chip-sequencing data using a deep learning framework

The identification of transcription factor binding sites and cis-regulatory motifs is a frontier whereupon the rules governing protein-DNA binding are being revealed. Here, we developed a new method (DEep Sequence and Shape mOtif or DESSO) for cis-regulatory motif prediction using deep neural networks and the binomial distribution model. DESSO outperformed existing tools, including DeepBind, in predicting motifs in 690 human ENCODE ChIP-sequencing datasets. Furthermore, the deep-learning framework of DESSO expanded motif discovery beyond the state-of-the-art by allowing the identification of known and new protein-protein-DNA tethering interactions in human transcription factors (TFs). Specifically, 61 putative tethering interactions were identified among the 100 TFs expressed in the K562 cell line. In this work, the power of DESSO was further expanded by integrating the detection of DNA shape features. We found that shape information has strong predictive power for TF-DNA binding and provides new putative shape motif information for human TFs. Thus, DESSO improves in the identification and structural analysis of TF binding sites, by integrating the complexities of DNA binding into a deep-learning framework

Development and Application of Software to Understand 3D Chromatin Structure and Gene Regulation

Nearly all cells contain the same 2 meters of DNA that must be systematically organized into their nucleus for timely access to genes in response to stimuli. Proteins and biomolecular condensates make this possible by dynamically shaping chromatin into 3D structures that connect regulators to their genes. Chromatin loops are structures that are partly responsible for forming these connections and can result in disease when disrupted or aberrantly formed. In this work, I describe three studies centered on using 3D chromatin structure to understand gene regulation. Using multi-omic data from a macrophage activation time course, we show that regulation temporally precedes gene expression and that chromatin loops play a key role in connecting enhancers to their target genes. In the next study, we investigated the role of biomolecular condensates in loop formation by mapping 3D chromatin structure in cell lines before and after disruption of NUP98-HOXA9 condensate formation. Differential analysis revealed evidence of CTCF-independent loop formation sensitive to condensate disruption. In the last study, we used 3D chromatin structure and multi-omic data in chondrocytes to link variant-gene pairs associated with Osteoarthritis (OA). Computational analysis suggests that a specific variant may disrupt transcription factor binding and misregulate inflammatory pathways in OA. To carry out these analyses I built computational pipelines and two R/Bioconductor packages to support the processing and analysis of genomic data. The nullranges package contains functions for performing covariate-matched subsampling to generate null-hypothesis genomic data and mitigate the effects of confounding. The mariner package is designed for working with large chromatin contact data. It extends existing Bioconductor tools to allow fast and efficient extraction and manipulation of chromatin interactions for better understanding 3D chromatin structure and its impact on gene regulation.Doctor of Philosoph

CRISPR-Cas: Development and applications for mammalian genome editing

The ability to introduce targeted modifications into genomes and engineer model organisms holds enormous promise for biomedical and technological applications, and has driven the development of tools such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). To facilitate genome engineering in mammalian cells, we have engineered the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 programmable nuclease systems from Streptococcus pyogenes SF370 (SpCas9) and S. thermophilus LMD-9 (St1Cas9) for mouse and human cell gene editing through heterologous expression of the minimal protein and RNA components. We have demonstrated that Cas9 nucleases can be guided by several short RNAs (sgRNAs) to introduce double stranded breaks (DSB) in the mammalian genome and induce efficient, multiplexed gene modification through non-homologous end-joining-mediated indels or homology-directed repair. Furthermore, we have engineered SpCas9 into a nicking enzyme (SpCas9n) to facilitate recombination while minimizing mutagenic DNA repair processes, and show that SpCas9n can be guided by pairs of appropriately offset sgRNAs to induce DSBs with high efficiency and specificity. In collaboration with Drs. Osamu Nureki and Hiroshi Nishimasu at the University of Tokyo, we further report the crystal structure of SpCas9 in complex with the sgRNA and target DNA, and elucidate the structure-function relationship of the ribonucleoprotein complex. Finally, through a metagenomic screen of orthologs, we have identified an additional small Cas9 from Staphylococcus aureus subsp. aureus (SaCas9) that cleaves mammalian endogenous DNA with high efficiency. SaCas9 can be packaged into adeno-associated virus for effective gene modification in vivo. Together, these technologies open up exciting possibilities for applications across basic science, biotechnology, and medicine

Carbohydrate-Active Enzymes

Carbohydrate-active enzymes are responsible for both biosynthesis and the breakdown of carbohydrates and glycoconjugates. They are involved in many metabolic pathways; in the biosynthesis and degradation of various biomolecules, such as bacterial exopolysaccharides, starch, cellulose and lignin; and in the glycosylation of proteins and lipids. Carbohydrate-active enzymes are classified into glycoside hydrolases, glycosyltransferases, polysaccharide lyases, carbohydrate esterases, and enzymes with auxiliary activities (CAZy database, www.cazy.org). Glycosyltransferases synthesize a huge variety of complex carbohydrates with different degrees of polymerization, moieties and branching. On the other hand, complex carbohydrate breakdown is carried out by glycoside hydrolases, polysaccharide lyases and carbohydrate esterases. Their interesting reactions have attracted the attention of researchers across scientific fields, ranging from basic research to biotechnology. Interest in carbohydrate-active enzymes is due not only to their ability to build and degrade biopolymers—which is highly relevant in biotechnology—but also because they are involved in bacterial biofilm formation, and in glycosylation of proteins and lipids, with important health implications. This book gathers new research results and reviews to broaden our understanding of carbohydrate-active enzymes, their mutants and their reaction products at the molecular level

56th Annual Rocky Mountain Conference on Magnetic Resonance

Final program, abstracts, and information about the 56th annual meeting of the Rocky Mountain Conference on Magnetic Resonance, co-endorsed by the Colorado Section of the American Chemical Society and the Society for Applied Spectroscopy. Held in Copper Mountain, Colorado, July 13-17, 2014

ICTERI 2020: ІКТ в освіті, дослідженнях та промислових застосуваннях. Інтеграція, гармонізація та передача знань 2020: Матеріали 16-ї Міжнародної конференції. Том II: Семінари. Харків, Україна, 06-10 жовтня 2020 р.

This volume represents the proceedings of the Workshops co-located with the 16th International Conference on ICT in Education, Research, and Industrial Applications, held in Kharkiv, Ukraine, in October 2020. It comprises 101 contributed papers that were carefully peer-reviewed and selected from 233 submissions for the five workshops: RMSEBT, TheRMIT, ITER, 3L-Person, CoSinE, MROL. The volume is structured in six parts, each presenting the contributions for a particular workshop. The topical scope of the volume is aligned with the thematic tracks of ICTERI 2020: (I) Advances in ICT Research; (II) Information Systems: Technology and Applications; (III) Academia/Industry ICT Cooperation; and (IV) ICT in Education.Цей збірник представляє матеріали семінарів, які були проведені в рамках 16-ї Міжнародної конференції з ІКТ в освіті, наукових дослідженнях та промислових застосуваннях, що відбулася в Харкові, Україна, у жовтні 2020 року. Він містить 101 доповідь, які були ретельно рецензовані та відібрані з 233 заявок на участь у п'яти воркшопах: RMSEBT, TheRMIT, ITER, 3L-Person, CoSinE, MROL. Збірник складається з шести частин, кожна з яких представляє матеріали для певного семінару. Тематична спрямованість збірника узгоджена з тематичними напрямками ICTERI 2020: (I) Досягнення в галузі досліджень ІКТ; (II) Інформаційні системи: Технології і застосування; (ІІІ) Співпраця в галузі ІКТ між академічними і промисловими колами; і (IV) ІКТ в освіті

Synthesis of new pyrazolium based tunable aryl alkyl ionic liquids and their use in removal of methylene blue from aqueous solution

In this study, two new pyrazolium based tunable aryl alkyl ionic liquids, 2-ethyl-1-(4-methylphenyl)-3,5- dimethylpyrazolium tetrafluoroborate (3a) and 1-(4-methylphenyl)-2-pentyl-3,5-dimethylpyrazolium tetrafluoroborate (3b), were synthesized via three-step reaction and characterized. The removal of methylene blue (MB) from aqueous solution has been investigated using the synthesized salts as an extractant and methylene chloride as a solvent. The obtained results show that MB was extracted from aqueous solution with high extraction efficiency up to 87 % at room temperature at the natural pH of MB solution. The influence of the alkyl chain length on the properties of the salts and their extraction efficiency of MB was investigated