Systematic screening of enhancer-blocking insulators in Drosophila identifies their DNA sequence determinants.

Long-range transcriptional activation of gene promoters by abundant enhancers in animal genomes calls for mechanisms to limit inappropriate regulation. DNA elements called insulators serve this purpose by shielding promoters from an enhancer when interposed. Unlike promoters and enhancers, insulators have not been systematically characterized due to lacking high-throughput screening assays, and questions regarding how insulators are distributed and encoded in the genome remain.

Protocol for detecting genomic insulators in Drosophila using insulator-seq, a massively parallel reporter assay.

Genomic insulators are DNA elements that prevent transcriptional activation of a promoter by an enhancer when interposed. We present a protocol for insulator-seq that enables high-throughput screening of genomic insulators using a plasmid-based massively parallel reporter assay in Drosophila cultured cells. We describe steps for insulator reporter plasmid library generation, transient transfection into cultured cells, and sequencing library preparation and provide a pipeline for data analysis.

Enhancer-promoter interactions become more instructive in the transition from cell-fate specification to tissue differentiation.

To regulate expression, enhancers must come in proximity to their target gene. However, the relationship between the timing of enhancer-promoter (E-P) proximity and activity remains unclear, with examples of uncoupled, anticorrelated and correlated interactions. To assess this, we selected 600 characterized enhancers or promoters with tissue-specific activity in Drosophila embryos and performed Capture-C in FACS-purified myogenic or neurogenic cells during specification and tissue differentiation.

Chromosome-level organization of the regulatory genome in the Drosophila nervous system.

Previous studies have identified topologically associating domains (TADs) as basic units of genome organization. We present evidence of a previously unreported level of genome folding, where distant TAD pairs, megabases apart, interact to form meta-domains. Within meta-domains, gene promoters and structural intergenic elements present in distant TADs are specifically paired.

Analysis of possible risk predictors in patients with coronavirus disease 2019: a retrospective cohort study.

Objective: This study aimed to analyze the clinical-epidemiological profile, possible risk predictors, and outcomes of patients with coronavirus disease 2019 admitted to the ward of a tertiary care hospital in southern Brazil. Specifically, we describe the demographic characteristics, comorbidities, baseline laboratory findings, clinical course, and survival of these patients.

Methods: This is an observational, retrospective cohort study, performed from January to March 2022, on medical records of patients hospitalized between April 2020 and December 2021 in the coronavirus disease 2019 ward of a tertiary hospital in southern Brazil.