AGO2 bound short RNAs in Jurkat cell line
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Project description:Characterization of AGO2 bound short RNAs from nuclear extracts of Jurkat cell line. Nuclei were isolated from cells; a fraction of Nuclear lysates was harvested and sequenced as the input samples. The rest of nuclear lysates were Immunoprecipitated using AGO2 monoclonal antibodies or Isotype matched IgG. Following immunoprecipitation, RNA was extracted from AGO2 or control IgG IP for short RNA sequencing. Two independent biological replicates were carried out.
2015-05-17 | E-MTAB-2104 | biostudies-arrayexpress
Project description:Characterization of AGO2 bound short RNAs from nuclear extracts of HeLaS3 cell line. Nuclei were isolated from HeLaS3 cells; a fraction of Nuclear lysates was harvsted and sequenced as the 'input' samples. The rest of nuclear lysates were Immunoprecipitated using AGO2 monoclonal antibodies or Isotype matched IgG. Following immunoprecipitation, RNA was extracted from AGO2 or control IgG IP for short RNA sequencing. Two independent biological replicates were carried out.
2015-05-17 | E-MTAB-2084 | biostudies-arrayexpress
Project description:Ago2-bound short RNAs in HeLaS3 cell line
| PRJEB4987 | ENA
Project description:Ago1-bound short RNAs in HeLaS3 cell line
| PRJEB5031 | ENA
Project description:Characterization of AGO1 bound short RNAs from nuclear extracts of HeLaS3 cell line. Nuclei were isolated from HeLaS3 cells; a fraction of Nuclear lysates was harvsted and sequenced as the input samples. The rest of nuclear lysates were immunoprecipitated using AGO1 monoclonal antibodies or Isotype matched IgG. Following immunoprecipitation, RNA was extracted from AGO1 or control IgG IP for short RNA sequencing. Two independent biological replicates were carried out.
2015-05-17 | E-MTAB-2129 | biostudies-arrayexpress
Project description:Among the four Argonaute family members in mammals, only AGO2 protein retains endonuclease activity, and facilitates cleavage of target RNAs base-pairing with highly complementary guide RNAs. Despite the deeply conserved catalytic activity, only a small number of targets have been reported to extensively base pair with cognate miRNAs to be cleaved by AGO2. Here, we analyzed AGO2-bound RNAs by CrossLinking ImmunoPrecipitation (CLIP) of genetically modified cells that express epitope-tagged AGO2 from the native genomic locus. We found that HMGA2 mRNA is cleaved by AGO2 loaded with let-7 and miR-21. In contrast to the generally accepted notion, the base-pairing from the seed region to the cleavage site, rather than perfect or near perfect complementarity, was required for cleavage of the target mRNA in cells. Non-templated addition of nucleotides at the 3’ end of the cleaved RNA was observed, further supporting the AGO2-mediated cleavage. Based on the observation that the limited complementarity is the minimum requirement for cleavage, we found that AGO2-mediated cleavage of targets is more common than previously thought. Our result may explain the vital role of endonuclease activity in controlling miRNA-mediated gene regulation.
2017-10-05 | GSE93644 | GEO
Project description:In order to identify miR-21 targets by a biochemical high-throughput method, we immunopurified RISC Complex and associated mRNAs in both control and miR-21 overexpressing Jurkat cells. Following transduction of miR-21 encoding construct (or a cognate control vector) in Jurkat cells, we used microarray technology to profile Total RNA, AGO2 Immunopurified RNA and control IgG purified RNA.
2014-10-15 | E-GEOD-37212 | biostudies-arrayexpress
Project description:Ago2-bound ShortRnas in HCT116 cell line and Dicer_hypomorphic(DICER_Ex5 cells) HCT116 cell line
| PRJEB4932 | ENA
Project description:Jurkat T cells have been exposed to 9g hypergravity in a custom-built pipette centrifuge for different times (GBF2021). Additionally, Jurkat T cells have been exposed to 300g for 5 minutes in a standard benchtop centrifuge, and waited for different times until adding lysis buffer. For comparison, Jurkat T cells have been exposed to 5 minutes of 42°C.
2023-02-07 | GSE221579 | GEO