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SRX20000327: GSM7181723: 1d030858; Zea mays; OTHER
1 ILLUMINA (Illumina NovaSeq 6000) run: 20.3M spots, 6.1G bases, 2Gb downloads

External Id: GSM7181723_r1
Submitted by: China Agricultural University
Study: Decoding the gene regulatory network of endosperm differentiation in maize [DAP-seq]
show Abstracthide Abstract
The persistent cereal endosperm constitutes the majority of the grain volume. Dissecting the gene regulatory network underlying cereal endosperm development will facilitate yield and quality improvement of cereal crops. Here, we use single-cell transcriptomics to analyze the developing maize (Zea mays) endosperm during cell differentiation. After obtaining transcriptomic data from 17,022 single cells, we identify 12 cell clusters corresponding to five endosperm cell types and revealing complex transcriptional heterogeneity. We delineate the temporal gene expression pattern from 6 to 7 days after pollination. We profile the genomic DNA binding sites of 161 transcription factors differentially expressed between cell clusters and constructed a gene regulatory network by combining the single-cell transcriptomic data with the direct DNA-binding profiles, identifying 181 regulons containing genes encoding transcription factors along with their high-confidence targets, Furthermore, we map the regulons to endosperm cell clusters, identify cell-cluster-specific essential regulators, and experimentally validated three predicted key regulators. This study provides a framework for understanding cereal endosperm development and function at single-cell resolution. Overall design: Construction of gene regulatory networks in maize endosperm using scRNA-seq and DAP-seq
Sample: 1d030858
SAMN34232837 • SRS17345045 • All experiments • All runs
Organism: Zea mays
Library:
Name: GSM7181723
Instrument: Illumina NovaSeq 6000
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: gDNA was extracted from maize B73 14 DAG leaf. gDNA libraries was prepared using TIANSeq DirectFast Library Kit (illumina) (cat. no. NG101, TIANGEN) according to the manufacturer's instructions. gDNA libraries was prepared using TIANSeq DirectFast Library Kit (illumina) (cat. no. NG101, TIANGEN) according to the manufacturer's instructions. Full length maize transcription factors' coding sequences were cloned from maize 14 DAG leaf cDNA into the NotI and AscI sites of pUC57-Halo using MultlF Seamless Assembly Mix (cat. no. RM20523, ABclonal). Protein expression was using TNT SP6 High-YIELD Wheat Germ Master Protein Expression System (cat. no. L3260; Promega) according to the manufacturer's instructions. Halo-fusion protein was bound to Magne HaloTag Beads (cat. no. G7281; Promega) and washed three times using TBSN buffer( 100 mM Tris buffer, pH 7.5, 50 mM NaCl, 0.005 % NP40). Then, 500 ng gDNA libraries was added and incubated at room temperature for 1.5 h. After incubation the beads was washed four times. Resuspended the beads in 30µl of EB buffer to recover DNA. The recovered DNA was were cleaned up with Silane magnetic beads, and amplified via PCR. The PCR products were cleaned up with Silane magnetic beads and sequenced using an NovaSeq platform (Illumina) by BerryGnomics.
Runs: 1 run, 20.3M spots, 6.1G bases, 2Gb
Run# of Spots# of BasesSizePublished
SRR2420366520,268,1506.1G2Gb2023-11-29

ID:
27393512

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