Targeted bisulfite sequencing in PTC
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1078879 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR28037900_1.fastq.gz | Fastqsanger.gz | |||
| SRR28037900_2.fastq.gz | Fastqsanger.gz | |||
| SRR28037901_1.fastq.gz | Fastqsanger.gz | |||
| SRR28037901_2.fastq.gz | Fastqsanger.gz | |||
| SRR28037902_1.fastq.gz | Fastqsanger.gz |
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Project description:Targeted bisulfite sequencing in PTC [ATAC-Seq]
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Project description:This study illuminates the critical advancement in the personalized management of papillary thyroid cancer (PTC) through the identification and validation of epigenetic biomarkers. We discovered the two distinct PTC subgroups with varying prognoses through TBS and observed the clues that PTC1 may related to immune signatures through multi-omics sequencing analysis. Then, we introduce FNAB, a minimally invasive preoperative test, for prognostication and therapeutic decision-making. Utilizing FNAB coupled with real-time digital PCR, our methodology not only facilitates the sensitive detection of seven pivotal biomarkers, including CD37, but also integrates seamlessly with the established TNM staging system to significantly refine survival predictions. This approach heralds a new era in the precise, personalized treatment of PTC, promising improved patient outcomes through the strategic application of epigenetic insights in clinical practice.
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Project description:This study illuminates the critical advancement in the personalized management of papillary thyroid cancer (PTC) through the identification and validation of epigenetic biomarkers. We discovered the two distinct PTC subgroups with varying prognoses through TBS and observed the clues that PTC1 may related to immune signatures through multi-omics sequencing analysis. Then, we introduce FNAB, a minimally invasive preoperative test, for prognostication and therapeutic decision-making. Utilizing FNAB coupled with real-time digital PCR, our methodology not only facilitates the sensitive detection of seven pivotal biomarkers, but also integrates seamlessly with the established TNM staging system to significantly refine survival predictions. This approach heralds a new era in the precise, personalized treatment of PTC, promising improved patient outcomes through the strategic application of epigenetic insights in clinical practice.
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Project description:We use NGS to assess the ability of TALE-guided DNA methyltranferases to make targeted changes to DNA methylation Targeted bisulfite sequencing of cells infected with wild-type or mutant TALE-DNMT constructs directed to the CDKN2A (p16) locus
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Project description:Cytosine methylation of DNA CpG dinucleotides in gene promoters is an epigenetic modification that regulates gene transcription. While many methods exist to interrogate methylation states, no current methods offer large-scale, targeted, single CpG resolution. We report an approach combining bisulfite treatment followed by RainDance microdroplet PCR with next-generation sequencing to assay the methylation state of 50 genes in the regions 1 kb upstream and downstream of their transcription start sites. Wildtype and hypermethylated Jurkat DNA (New Englad Biolabs) was treated with bisulfite to convert all unmethylated cytosines to uracil. Following bisulfite treatment, targeted amplification was carried out using a custom primer library and microdroplet PCR. PCR product was sheared to 200 bp and ligated to sequencing adapters following standard protocols. Sequencing was conducted with single-end 100 bp reads on an Illumina GAIIx for wild type Jurkat DNA or Jurkat CpG DNA with a single sample per lane.
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Project description:We report a method for specific capture of an arbitrary subset of genomic targets for single molecule bisulfite sequencing, and for digital quantitation of DNA methylation at a single nucleotide resolution. We used targeted bisulfite sequencing to characterize the changes of DNA methylation during the de-differentiation of human fibroblasts into hybrid stem cells, and into induced pluripotent stem cells. We compared the methylation level of approximately 66,000 CpG sites within 2020 CpG islands on chromosome 12, chromosome 20, and 34 selected regions. A total of 288 differentially methylated regions were identified between fibroblasts and pluripotent cells. Methylation cluster analysis revealed distinct methylation patterns between fibroblasts and pluripotent cells. Furthermore iPS cells are globally more methylated than human embryonic stem cells, which could be due to the reprogramming process. This targeted bisulfite sequencing method is particularly useful for efficient and large-scale analysis of DNA methylation in organisms with large genomes. Experiment Overall Design: Comparison of DNA methylation on 2020 CpG islands and 34 other selected regions among eleven human ES, iPS and fibroblast lines.
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Project description:In vertebrates, DNA methylation-mediated repression of retrotransposons is essential for the maintenance of genomic integrity. In the current study, we developed a technique termed HT-TREBS (High-Throughput Targeted Repeat Element Bisulfite Sequencing). This technique is designed to measure the DNA methylation levels of individual loci of any repeat families with next-generation sequencing approaches. To test the feasibility of HT-TREBS, we analyzed the DNA methylation levels of the IAPLTR family using a set of 12 different genomic DNA isolated from the brain, liver and kidney of 4 one-week-old littermates of the mouse strain C57BL/6N. This technique has successfully generated the CpG methylation data of 5,233 loci common in all the samples, representing more than 80% of the individual loci of the five targeted subtypes of the IAPLTR family. According to the results, approximately 5% of the IAPLTR loci have less than 80% average CpG methylation levels with no genomic position preference. Further analyses of the IAPLTR loci also revealed the presence of extensive DNA methylation variations between different tissues and individuals. Overall, these data demonstrate the efficiency and robustness of the new technique, HT-TREBS, and also provide new insights regarding the genome-wide DNA methylation patterns of the IAPLTR repeat elements. High-throughput, single-base resolution, singlicate DNA methylation profiles of IAPLTR retrotransposons in the brain, liver , and kidney of four 1-week-old mouse littemates using the developed technique, HT-TREBS.
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