Publications by authors named "Robert Querfurth"

A substantial fraction of phenotypic differences between closely related species are likely caused by differences in gene regulation. While this has already been postulated over 30 years ago, only few examples of evolutionary changes in gene regulation have been verified. Here, we identified and investigated binding sites of the transcription factor GA-binding protein alpha (GABPa) aiming to discover cis-regulatory adaptations on the human lineage.

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Since the introduction of next-generation sequencing, several techniques have been developed to selectively enrich and sequence specific parts of the genome at high coverage. These techniques include enzymatic methods employing molecular inversion probes, PCR based approaches, hybrid capture, and in-solution capture. In-solution capture employs RNA probes transcribed from a pool of DNA template oligos designed to match regions of interest to specifically bind and enrich genomic DNA fragments.

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Background: The interplay between transcription and mutational processes can lead to particular mutation patterns in transcribed regions of the genome. Transcription introduces several biases in mutational patterns; in particular it invokes strand specific mutations. In order to understand the forces that have shaped transcripts during evolution, one has to study mutation patterns associated with transcription across animals.

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Article Synopsis
  • The study emphasizes that experimental validation is essential for understanding mammalian promoters and cis-regulatory elements, as DNA sequence analysis alone isn't enough.
  • Researchers focused on human chromosome 21, cloning 182 gene promoters and performing assays, discovering 56 active promoters in HEK293 cells and an additional 49 that could be activated under certain conditions.
  • Findings showed that while truncating many promoters to 500 bp generally maintained activity, it diminished their responsiveness to external stimuli, hinting at important regulatory elements located farther upstream.
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Article Synopsis
  • Promoters are key transcriptional regulatory sequences located upstream of gene start sites, but identifying all human transcription start sites and their functional elements remains incomplete.
  • A study examined 74 promoters related to inflammatory bowel disease using a transfected-cell array in HEK293T cells, revealing that 21.6% were active.
  • The study found that 75% of these active promoters corresponded with their gene's transcriptional activity, highlighting the importance of promoter activity in gene expression regulation and showcasing a novel large-scale approach to studying regulatory sequences.
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Polymerase chain reaction (PCR) has become a fundamental technique in molecular biology. Nonetheless, further improvements of the existing protocols are required to broaden the applicability of PCR for routine diagnostic purposes, to enhance the specificity and the yield of PCRs as well as to reduce the costs for high-throughput applications. One known problem typically reported in PCR experiments is the poor amplification of GC-rich DNA sequences.

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