Basic Information

Dataset GSE192376
Status 2022-05-18
Species Homo sapiens
Source NCBI GEO
Subcellular Localization nuclear|cytosolic
PMID 35589130
Title RNA-fractionation sequencing
Citation Thomas A, Rehfeld F, Zhang H, Chang TC et al. RBM33 directs the nuclear export of transcripts containing GC-rich elements. Genes Dev 2022 May 1;36(9-10):550-565.
Experimental Design 400 000 WT, RBM33 KO-1, or RBM33 KO-2 HCT116 cells were seeded in 6-well plates. For each replicate, 48 hours later, one well was subjected to total RNA extraction and one well to nuclear-cytoplasmic fractionationFractionation was performed as described previously (Lee et al., 2016). Briefly, cells were collected by trypsinization and lysed in 175 µL RLN1 buffer [50 mM Tris-HCl pH 8, 140 mM NaCl, 1.5 mM MgCl2, 0.5% NP-40, and 1:1000 SUPERase In RNase inhibitor (Thermo Fisher Scientific)]. Following a 5-minute incubation on ice, the sample was centrifuged at 300 g for 2 minutes. The supernatant, representing the cytoplasmic fraction, was collected and centrifuged at 16000 g for 5 minutes to clear debris. The initial pellet, representing the nuclear fraction, was gently resuspended in 500 µL RLN1 and centrifuged at 1000 g for 5 minutes to wash. The pellet was then resuspended in 175 µL RLN2 buffer [50 mM Tris-HCl pH 8, 500 mM NaCl, 1.5 mM MgCl2, 0.5% NP-40, and 1:1000 SUPERase In RNase inhibitor (Thermo Fisher Scientific)]. RNA was extracted from each of these fractions by Trizol in combination with the miRNeasy mini kit (Qiagen) with an on-column DNase digestion.

Sample Information

BioProject Sample Name Localization Genotype Source Label
PRJNA791311 GSM5745792 cytosolic RNA fraction RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 1 cytosolic fraction
PRJNA791311 GSM5745793 nuclear RNA fraction RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 1 nuclear fraction
PRJNA791311 GSM5745794 total RNA RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 1 total
PRJNA791311 GSM5745795 cytosolic RNA fraction RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 2 cytosolic fraction
PRJNA791311 GSM5745796 nuclear RNA fraction RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 2 nuclear fraction
PRJNA791311 GSM5745797 total RNA RBM33 KO clone1 HCT116 colon cancer cell line RBM33 KO clone1 replicate 2 total
PRJNA791311 GSM5745798 cytosolic RNA fraction RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 1 cytosolic fraction
PRJNA791311 GSM5745799 nuclear RNA fraction RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 1 nuclear fraction
PRJNA791311 GSM5745800 total RNA RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 1 total
PRJNA791311 GSM5745801 cytosolic RNA fraction RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 2 cytosolic fraction
PRJNA791311 GSM5745802 nuclear RNA fraction RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 2 nuclear fraction
PRJNA791311 GSM5745803 total RNA RBM33 KO clone2 HCT116 colon cancer cell line RBM33 KO clone2 replicate 2 total
PRJNA791311 GSM5745804 cytosolic RNA fraction WT HCT116 colon cancer cell line WT replicate 1 cytosolic fraction
PRJNA791311 GSM5745805 nuclear RNA fraction WT HCT116 colon cancer cell line WT replicate 1 nuclear fraction
PRJNA791311 GSM5745806 total RNA WT HCT116 colon cancer cell line WT replicate 1 total
PRJNA791311 GSM5745807 cytosolic RNA fraction WT HCT116 colon cancer cell line WT replicate 2 cytosolic fraction
PRJNA791311 GSM5745808 nuclear RNA fraction WT HCT116 colon cancer cell line WT replicate 2 nuclear fraction
PRJNA791311 GSM5745809 total RNA WT HCT116 colon cancer cell line WT replicate 2 total
PRJNA791311 GSM5745810 cytosolic RNA fraction WT HCT116 colon cancer cell line WT replicate 3 cytosolic fraction
PRJNA791311 GSM5745811 nuclear RNA fraction WT HCT116 colon cancer cell line WT replicate 3 nuclear fraction
PRJNA791311 GSM5745812 total RNA WT HCT116 colon cancer cell line WT replicate 3 total

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