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Series GSE196427 Query DataSets for GSE196427
Status Public on Mar 22, 2022
Title The XRN1-regulated RNA helicase activity of YTHDC2 ensures mouse fertility independently of m6A recognition
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Other
Summary The functional consequence of N6-methyladenosine (m6A) RNA modification is mediated by ‘reader’ proteins like those of the YTH family that recruit additional factors to alter splicing, RNA stability or translation. Germline-specific YTH domain-containing 2 (YTHDC2) is essential for mammalian fertility, but its molecular function is poorly understood. Here we use a tagged mouse line to identify U-rich motifs as binding sites of YTHDC2 on 3′ UTRs of testicular RNA targets. Although its YTH domain is an m6A-binder in vitro, the YTH point mutant mouse lacking this ability displays no obvious phenotypes. Its 3′→5′ RNA helicase activity is essential for fertility, with the catalytic-dead mutation being dominant-negative. Single-cell transcriptomics reveal that Ythdc2 mutant mitotic testicular germ cells transition into meiosis but accumulate a transcriptome with mixed mitotic/meiotic identity. We also reveal a conserved role for YTHDC2 in animal fertility.
 
Overall design iCLIP experiment was used to identify the RNA targets of YTHDC2, as well as its binding sites and motifs. RNA sequencing was applied to study the gene expression differences between Ythdc2YTHmut/+ and Ythdc2YTHmut/YTHmut of testicular 2n and 4n cells, Ythdc2+/+, Ythdc2+/cat-dead, and Ythdc2cat-dead/cat-dead.
 
Contributor(s) Li L, Krasnykov K, Homolka D, Gos P, Mendel M, Fish R, Raman Pandey R, Pillai RS
Citation(s) 35305312
Submission date Feb 09, 2022
Last update date Mar 22, 2022
Contact name Ramesh Pillai
E-mail(s) ramesh.pillai@unige.ch
Organization name University of Geneva
Department Department of Molecular Biology
Street address 30, Quai Ernest-Ansermet
City Gneveva
ZIP/Postal code CH-1211
Country Switzerland
 
Platforms (2)
GPL19057 Illumina NextSeq 500 (Mus musculus)
GPL21103 Illumina HiSeq 4000 (Mus musculus)
Samples (25)
GSM5883825 Ythdc2_WT_P10_embryo [RR796; scRNA]
GSM5883826 Ythdc2_KO1_P10_embryo [RR797; scRNA]
GSM5883827 Ythdc2_KO_P10_embryo [RR798; scRNA]
Relations
BioProject PRJNA804877

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE196427_YTH_mut_DE_tables.xlsx 10.0 Mb (ftp)(http) XLSX
GSE196427_Ythdc2_cat_dead_DE_tables.xlsx 6.4 Mb (ftp)(http) XLSX
GSE196427_Ythdc2_scRNA-seq_cells.csv.gz 410.6 Kb (ftp)(http) CSV
GSE196427_Ythdc2_scRNA-seq_genes.csv.gz 536.7 Kb (ftp)(http) CSV
GSE196427_Ythdc2_scRNA-seq_matrix.csv.gz 92.9 Mb (ftp)(http) CSV
GSE196427_iCLIP_final_tab_per_gene.xlsx 1.8 Mb (ftp)(http) XLSX
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Raw data are available in SRA
Processed data are available on Series record

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