GSE196331
Reducedfemalefertility duetosequestration ofRNA Pol II by pervasive transcription in exosome RNasedepleted oocytes
We performed mouse single oocyte RNA-seq and bulk oocyte CUT&Tag assays in the current project. In details, SMART-seq based single oocyte RNA-seq was performed at adult GV stage, GV3h stage and MII stage, using control and Dis3 oocyte-speicfic knockout (cKO) oocytes. RiboMinus-seq based single oocyte RNA-seq was performed at adult GV stage using control and Dis3 cKO oocytes, and at p20 GV stage using control, Dis3 cKO, Exosc10 cKO and Dis3/Exosc10 double cKO (dcKO) oocytes. Bulk CUT&Tag of anti-H3K27me3 was done in WT and Dis3 cKO oocytes at adult GV stage, and in WT and Dis3/Exosc10  dcKO oocytes at p20 stage. In addition, CUT&Tag of anti-RNA polymerase II (Ser2+Ser5) was performed in WT and Dis3 cKO oocytes at GV stage. All CUT&Tag experiments share the same rabbit-Igg negative control. All p20 stage oocytes were specified as p20. The non-specified GV oocytes were all adult GV oocytes. Overall design: In SMART-seq based single oocyte RNA-seq, totally 72 samples were performed, including 24 samples at GV stage, 24 samples at GV3h stage and 24 samples at MII stage. At each stage, 8 samples were from control group and 16 samples were from Dis3 cKO group. In RiboMinus RNA-seq, all groups (adult-GV-ctr, adult-GV-Dis3cKO, p20-ctr, p20-Dis3cKO, p20-Exosc10, p20-dcKO) have 8 samples each. In CUT&Tag assays, each group has 3 replicates in totall. The CUT&Tag sequencing results were all firstly randomly sampled for 3M reads for downstream analysis (without deduplication).