{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"submitter":["Oscar Juez"],"organism":["Arabidopsis thaliana"],"software":["base calling and demultiplexing"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16251"],"description":["RNA-seq experiment to characterize transcriptome changes associated with premature aging in Arabidopsis nuclear-envelope mutants. Col-0 wild type and single mutants in CRWN1, CRWN2, CRWN3, CRWN4 and KAKU4 were grown on soil under short-day conditions (8 h light / 16 h dark) until 150 days after sowing, a stage at which nuclear-envelope mutants display strong rosette senescence and loss of meristem activity. Entire rosettes were harvested, immediately frozen, and pooled per genotype to capture whole-plant vegetative transcriptional states. Total RNA was extracted from rosette tissue, mRNA was enriched, and strand-specific Illumina libraries were prepared and sequenced as paired-end short reads. The resulting dataset is intended to (i) compare global gene-expression profiles between wild type and individual nuclear-envelope mutants at late vegetative stages, (ii) identify pathways associated with progeroid-like phenotypes, and (iii) provide a resource for linking nuclear architecture defects to age-related transcriptional drift in plants."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Sample Collection - Arabidopsis thaliana Col-0 background plants (wild type, crwn1–4, kaku4) were grown under short-day conditions and sampled at 150 days after sowing. From each rosette, two defined leaf cohorts were dissected: • Basal pool: leaves 1–4 (oldest fully expanded rosette leaves) • Apical pool: leaves 11–14 (most recently formed photosynthetic leaves) Basal and apical pools from the same plant were combined to represent whole-rosette profiles, immediately flash-frozen in liquid nitrogen, and stored at −80 °C. Each biological replicate consisted of pooled tissue from 3–4 plants (one pot), and two biological replicates per genotype were collected.","Growth Protocol - Arabidopsis thaliana Col-0 wild type and nuclear-envelope single mutants (crwn1–4, kaku4) were grown in soil in a controlled-environment growth chamber under short-day photoperiod (8 h light / 16 h dark) at 22 °C with ~60 % relative humidity. Light intensity was maintained at approximately 120–150 µmol m⁻² s⁻¹ of white light. Plants were bottom-watered regularly and fertilized according to standard greenhouse practice. Three pots (3–4 plants per pot) per genotype were established; two/three pots per genotype were used for molecular profiling, and one pot was maintained as a backup under identical conditions.","Nucleic Acid Extraction - Frozen pooled rosette tissue was ground to a fine powder in liquid nitrogen. Total RNA was extracted using the RNeasy Plant Mini Kit (Qiagen, cat. 74904) following the manufacturer’s instructions, including on-column DNase digestion to remove genomic DNA. RNA concentration was measured by Qubit RNA HS assay and purity was assessed by A260/280 and A260/230 ratios. RNA integrity was evaluated on an Agilent Bioanalyzer, and only samples with sufficient yield and high RIN scores were used for RNA-seq library preparation.","Sequencing - Indexed libraries were pooled equimolarly and sequenced at the OIST Sequencing Section (SQC) on an Illumina NovaSeq 6000 platform using paired-end 2 × 150 bp reads. Standard Illumina base calling and demultiplexing were used to generate gzipped FASTQ files per sample. No additional in-run spike-ins were used beyond standard dual indices.","Library Construction - For each sample, 100 ng of high-quality total RNA was depleted of ribosomal RNA using QIAseq FastSelect reagents according to the manufacturer’s protocol. rRNA-depleted RNA was used to prepare strand-specific libraries with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB, E7760). Briefly, rRNA-depleted RNA was fragmented at 94 °C for 8 min, followed by first-strand cDNA synthesis (25 °C 10 min, 42 °C 50 min, 70 °C 15 min) and dUTP-based second-strand synthesis per kit instructions. After SPRIselect 1.8× cleanup, end repair, A-tailing, adaptor ligation, USER treatment (37 °C, 15 min), and a 0.9× cleanup, libraries were amplified with 12 PCR cycles using NEBNext Ultra II Q5 polymerase. Final libraries were bead-purified, size-checked on a Bioanalyzer (High Sensitivity or DNA 1000), and quantified by Qubit dsDNA HS assay."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"omics_type":["Unknown","Transcriptomics","Genomics","Proteomics"],"instrument_platform":["N/A","Thermocycler; magnetic rack for SPRI bead clean-up","Illumina NovaSeq 6000","Liquid-nitrogen grinding tools; benchtop centrifuge","Controlled-environment growth chamber (short-day; 8 h light / 16 h dark)"],"study_type":["RNA-seq of total RNA"],"species":["Arabidopsis thaliana"],"pubmed_authors":["Oscar Juez"],"additional_accession":[]},"is_claimable":false,"name":"RNA-seq of Arabidopsis thaliana Col-0 wildtype and single mutants crwn1, crwn2, crwn3, crwn4 and kaku4 in short-day conditions collected at 150 days after sowing","description":"RNA-seq experiment to characterize transcriptome changes associated with premature aging in Arabidopsis nuclear-envelope mutants. Col-0 wild type and single mutants in CRWN1, CRWN2, CRWN3, CRWN4 and KAKU4 were grown on soil under short-day conditions (8 h light / 16 h dark) until 150 days after sowing, a stage at which nuclear-envelope mutants display strong rosette senescence and loss of meristem activity. Entire rosettes were harvested, immediately frozen, and pooled per genotype to capture whole-plant vegetative transcriptional states. Total RNA was extracted from rosette tissue, mRNA was enriched, and strand-specific Illumina libraries were prepared and sequenced as paired-end short reads. The resulting dataset is intended to (i) compare global gene-expression profiles between wild type and individual nuclear-envelope mutants at late vegetative stages, (ii) identify pathways associated with progeroid-like phenotypes, and (iii) provide a resource for linking nuclear architecture defects to age-related transcriptional drift in plants.","dates":{"release":"2026-06-22T00:00:00Z","modification":"2026-06-22T01:00:38.083Z","creation":"2025-11-22T22:06:44.769Z"},"accession":"E-MTAB-16251","cross_references":{"ENA":["ERP185510"],"EFO":["EFO_0002944","EFO_0004170","EFO_0009653","EFO_0003789","EFO_0005518","EFO_0004184"]}}