Structural and genetic characterization of chromosomes is necessary to understand both normal and pathologic physiology in any species. Flow cytometry and cell sorting technologies provide a means for precise measurement of chromosomal makeup as well as for the isolation of specific chromosomes for further study. Advancements in molecular biology protocols and pressures from large-scale sequencing endeavors placed increased demand on the developers of these instruments for enhanced throughput and quality of results. The ensuing improvements in sorting performance led to the development of a new generation of cytometers known as high-speed cell sorters. These machines provide superior results in less time and are cheaper and simpler to operate than their predecessors. Robust chromosome sorting can now be performed in the laboratories of individual investigators for a variety of gene- and sequence-specific studies. Resolution of the flow karyotype with increased refinement, and the development of new applications for this technology will assure that cell sorting continues to play an important role in cytogenetics, our understanding of molecular processes such as evolution and disease etiology, and ultimately serve as a launching point for predictive medicine.
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http://dx.doi.org/10.1023/b:chro.0000009328.96958.a6 | DOI Listing |
Nat Commun
January 2025
IBENS, Département de biologie, École normale supérieure, Université PSL, CNRS, INSERM, 75005, Paris, France.
Current temporal studies of DNA replication are either low-resolution or require complex cell synchronisation and/or sorting procedures. Here we introduce Nanotiming, a single-molecule, nanopore sequencing-based method producing high-resolution, telomere-to-telomere replication timing (RT) profiles of eukaryotic genomes by interrogating changes in intracellular dTTP concentration during S phase through competition with its analogue bromodeoxyuridine triphosphate (BrdUTP) for incorporation into replicating DNA. This solely demands the labelling of asynchronously growing cells with an innocuous dose of BrdU during one doubling time followed by BrdU quantification along nanopore reads.
View Article and Find Full Text PDFSci Rep
December 2024
Division of Genetics, Indian Agricultural Research Institute, New Delhi, 110012, India.
The mungbean yellow mosaic India virus (MYMIV, Begomovirus vignaradiataindiaense) causes Yellow Mosaic Disease (YMD) in mungbean (Vigna radiata L.). The biochemical assays including total phenol content (TPC), total flavonoid content (TFC), ascorbic acid (AA), DPPH (2,2-diphenyl-1-picrylhydrazyl), and FRAP (Ferric Reducing Antioxidant Power) were used to study the mungbean plants defense response to MYMIV infection.
View Article and Find Full Text PDFJ Hered
December 2024
Ohio Biodiversity Conservation Partnership, The Ohio State University, Columbus, Ohio, 43210 USA.
Conservation of threatened species can benefit from an evaluation of genes in the Major Histocompatibility Complex (MHC), whose loci encode proteins that bind pathogens and are often under strong selection to maintain diversity in immune response to diseases. Despite this gene family's importance to disease resistance, little is known about these genes in reptiles including snakes. To address this issue, we assembled and annotated a highly-contiguous genome assembly for the timber rattlesnake (Crotalus horridus), a pit viper which is threatened or endangered in parts of its range, and analyzed this new genome along with three other rattlesnake genomes to characterize snake MHC loci.
View Article and Find Full Text PDFCell Rep Methods
December 2024
Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Genetics and Genomic Sciences, Center for Advanced Genomics Technology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address:
We apply a single-molecule chromatin fiber sequencing (Fiber-seq) protocol designed for amplification-free cell-type-specific mapping of the regulatory architecture at nucleosome resolution along extended ∼10-kb chromatin fibers to neuronal and non-neuronal nuclei sorted from human brain tissue. Specifically, application of this method enables the resolution of cell-selective promoter and enhancer architectures on single fibers, including transcription factor footprinting and position mapping, with sequence-specific fixation of nucleosome arrays flanking transcription start sites and regulatory motifs. We uncover haplotype-specific chromatin patterns, multiple regulatory elements cis-aligned on individual fibers, and accessible chromatin at 20,000 unique sites encompassing retrotransposons and other repeat sequences hitherto "unmappable" by short-read epigenomic sequencing.
View Article and Find Full Text PDFGenetics
November 2024
Department of Biological Sciences, University of Denver, Denver, CO 80208, USA.
Hybrid incompatibilities are a critical component of species barriers and may arise due to negative interactions between divergent regulatory elements in parental species. We used a comparative approach to identify common themes in the regulatory phenotypes associated with hybrid male sterility in two divergent rodent crosses, dwarf hamsters and house mice. We investigated three potential characteristic gene expression phenotypes in hybrids including the propensity of transgressive differentially expressed genes towards over or underexpression, the influence of developmental stage on patterns of misexpression, and the role of the sex chromosomes on misexpression phenotypes.
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