Scientific Posters

Zymo Research's scientific posters highlight the latest breakthroughs in molecular biology, showcasing innovations spanning from epigenetics to microbiomics. We are proud to present insights from our collaborations with leading scientists and institutions worldwide, with more contributions to the field of life science on the horizon.

Epigenetic Biomarker Discovery and Validation for Diagnosis and Therapeutic Intervention for Hepatocellular Carcinoma
Xueguang Sun, Jill E. Petrisko, Lam K. Nguyen, Marc Van Eden & Xi-Yu Jia

Hepatocellular carcinoma (HCC) is one of the most common and lethal malignancies worldwide, accounting for approximately half a million annual deaths globally. HCC is completely asymptomatic in the early stages of the disease; therefore, early detection of HCC in afflicted patients is vital to receive therapeutic benefits from curative surgery. The standard diagnosis of HCC relies upon detection of the serum alpha-fetoprotein (AFP) level in at-risk subjects followed by hepatic ultrasonography to identify suspicious nodules. Accurate levels of AFP are often difficult to detect, and the imaging method to identify nodules is both operator-sensitive and subject to a high false-negative rate. While it is known that hepatocellular carcinoma is a multi-step process that requires altered expression of multiple genes, recent evidence has indicated that epigenetic abnormalities also play an important role in HCC. The discovery of reliable and accurate epigenetic biomarkers may open up new avenues for the development of novel diagnostic tools and provide a new target for therapeutic interventions.

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EpiQuest- A Genomic Approach for DNA Methylation and Hydroxymethylation Analysis
Xueguang Sun, TzuHung Chung, Eliza Bacon, Ron Leavitt, Nikolas Isely, Marc Van Eden & Xi-Yu Jia

DNA methylation and hydroxymethylation are some of the most important epigenetic modifications that can occur to the human genome. For instances, DNA methylation plays a vital role in the regulation of gene expression in normal cell development and aging, but also in the formation and progression of cancer and other diseases. Profiling DNA methylation and hydroxymethylation at the genome level promises large-scale identification of epigenetic biomarkers that could be applied to clinical and molecular diagnostic fields. Due to the availability of Next Gen sequencing technology, a number of new technologies have been developed for interrogating DNA methylation and hydroxymethylation at the genomic scale. Zymo Research has recently perfected sample prep and bioinformatic analysis as part of its new EpiQuest™ Methylation and Hydroxymethylation Profiling Services. These services combine next generation sequencing with Zymo's wellestablished epigenetic technologies and innovative bioinformatic algorithms for the most streamlined, comprehensive genome scale data generation to date. With EpiQuest… hundreds of epigenomic biomarker candidates can be discovered at once.

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Genome-Wide DNA Methylation Analysis in Autism
Xi-Yu Jia, Xueguang Sun, Eliza Bacon, Adam Peterson, TzuHung Chung

Lacking consistent genetic mutation data in autism and increased risk of prenatal /maternal factors for disease development suggest a possible epigenetic mechanism for the disease development. DNA methylation is one of the major epigenetic regulators and its importance in development and disease is well established. Using next generation sequencing in combination with bisulfite-based DNA methylation detection, genome-wide 5-methylcytosine (5-mC) were investigated in autism blood samples from monozygotic twins. Our results indicate that “epimutations” are present in the affected children's blood DNA and these epigenetic changes were in agreement with other published biochemical data implicating epigenetics as a key element in the development of the disease.

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Genome-Wide DNA Methylation Analysis In Plants and Animals
Xueguang Sun, TzuHung Chung, Eliza Bacon, Ron Leavitt, Nikolas Isely, Marc Van Eden & Xi-Yu Jia

DNA methylation is a highly conserved epigenetic mark present in many eukaryotic organisms including plants, animals, and fungi. It plays an important role in the regulation of gene expression. A number of studies have shown its involvement in plant and animal growth and reproduction primary through the processes of genomic imprinting, X-chromsome inactivation, and the silencing of transposons and other repetitive DNA elements. As such, the understanding of DNA methylation has become a major focus of the research conducted during the “post-genomics” era. However, the precise determination of a DNA's methylation pattern on a genomic scale has posed a challenge especially for those complex genomes. Combining well-established bisulfite conversion chemistry with NextGen sequencing, we have established a robust service platform for analyzing DNA methylation with single base resolution at the genomic scale. The service features a streamlined workflow coupled with a comprehensive bioinformatics pipeline to provide both a consolidated and cost-effective solution for epigenetic analysis of plant and animal genomes. This technology has been used successfully for the analysis of methylomes from many organisms including soybean, mouse, and chicken. Additional analyses of other species are ongoing. Those data should provide a means to understanding how environment as well as other factors may alter an organism's fitness through heritable changes in epigenetic gene expression.

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Genome-Wide Human Brain DNA 5-hmC Profiling Using a Novel Sequence- and Strand-Specific Method
Xueguang Sun, Adam Petterson, Tzu Hung Chung, Xi Yu Jia, Pu Zhang

5-Hydroxymethylcytosine (5-hmC) is an epigenetic hallmark which has recently become central in mapping and sequencing work. While the exact function of this base is not fully understood, it is likely to regulate gene expression as a member of active DNA demethylation pathways. The levels of 5-hmC in genomic DNA vary significantly depending on the cell type, though the highest levels are found in cells of the central nervous system (CNS): These findings suggest importance of 5- hmC in gene regulation within the CNS. While several methods have been developed to profile 5-hmC at genomic scale, most are enrichment-based, utilize large amounts of genomic DNA input, and have relatively low resolution. Although efforts have been made to detect 5hmC at single-site resolution, the methods described to date still require several micrograms of DNA, require parallel or subtractive sequencing, and employ successive chemical treatments that degrade the DNA and hinder sequencing. By combining modification-sensitive restriction enzymes with massively parallel (“next-generation”) sequencing approaches, we developed a novel Reduced Representation Hydroxymethylation Profiling (RRHP) method for genome-wide 5-hmC mapping at single-site resolution from low (100 ng) DNA inputs. Importantly, the method can detect strand polarity of 5-hmC modifications, and also enables the direct identification of single nucleotide polymorphisms (SNPs) within sequencing reads. Due to the fragmentation approach, data can be directly compared with single-base DNA methylation data from Reduced Representation Bisulfite Sequencing (RRBS). Human brain 5-hmC mapping generated with this method, combined with DNA methylation profiling data, indicates unique distributions of 5-hmC modification: We confirm that several important neuronal loci, such as BDNF, NLGN2, CES1, and TAF1, demonstrate extensive 5-hmC modification. This new method of detection and mapping is a powerful tool in enhancing our understanding of the interplay of genetic and epigenetic regulations in neurobiology and other diverse biological fields

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