Mus musculus
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA175440 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Recurrent somatic hotspot mutations of DICER1 appear to be clustered around each of four critical metal binding residues in the RNase IIIB domain of DICER1. This domain is responsible for cleavage of the 3’ end of the 5p-miRNA strand of a pre-mRNA hairpin. To investigate the effects of these cancer-associated “hotspot” mutations we engineered mouse Dicer1-deficient ES cells to express wild-type and an allelic series of the mutant human DICER1 variants. Global miRNA and mRNA profiles from cells carrying the metal binding site mutations were compared to each other and wild-type human DICER1. The miRNA and mRNA profiles generated through the expression of the hotspot mutations were virtually identical, and the DICER1 hotspot mutation carrying cells were distinct from both wild-type and Dicer1-deficient cells. Further, miRNA profiles showed mutant DICER1 results in a dramatic loss in processing of mature 5p-miRNA strands but were still able to create 3p-strand miRNAs. Messenger-RNA profile changes were consistent with the loss of 5p-strand miRNAs and showed enriched expression for predicted targets of the lost 5p derived miRNAs. We therefore conclude that cancer-associated somatic hotspot mutations of DICER1, affecting any one of four metal binding residues in the RNase IIIB domain, are functionally equivalent with respect to miRNA-processing and are hypomorphic alleles, yielding a global loss in processing of mature 5p-strand miRNA. We further propose that this resulting 3p-strand bias in mature miRNA expression likely underpins the oncogenic potential of these hotspot mutations. A total of 28 Affymetrix Mouse Gene ST arrays were done for mRNA expression profiling of various DICER1 mutants (n=14), wildtype controls (n=6), vector only (n=3) and parental cell lines (n=5).
2012-10-01 | E-GEOD-40965 | biostudies-arrayexpress
Project description:Recurrent somatic hotspot mutations of DICER1 appear to be clustered around each of four critical metal binding residues in the RNase IIIB domain of DICER1. This domain is responsible for cleavage of the 3’ end of the 5p-miRNA strand of a pre-mRNA hairpin. To investigate the effects of these cancer-associated “hotspot” mutations we engineered mouse Dicer1-deficient ES cells to express wild-type and an allelic series of the mutant human DICER1 variants. Global miRNA and mRNA profiles from cells carrying the metal binding site mutations were compared to each other and wild-type human DICER1. The miRNA and mRNA profiles generated through the expression of the hotspot mutations were virtually identical, and the DICER1 hotspot mutation carrying cells were distinct from both wild-type and Dicer1-deficient cells. Further, miRNA profiles showed mutant DICER1 results in a dramatic loss in processing of mature 5p-miRNA strands but were still able to create 3p-strand miRNAs. Messenger-RNA profile changes were consistent with the loss of 5p-strand miRNAs and showed enriched expression for predicted targets of the lost 5p derived miRNAs. We therefore conclude that cancer-associated somatic hotspot mutations of DICER1, affecting any one of four metal binding residues in the RNase IIIB domain, are functionally equivalent with respect to miRNA-processing and are hypomorphic alleles, yielding a global loss in processing of mature 5p-strand miRNA. We further propose that this resulting 3p-strand bias in mature miRNA expression likely underpins the oncogenic potential of these hotspot mutations.
2012-10-01 | GSE40965 | GEO
Project description:The DICER1 gene is mutated in cancer, including Dicer1 syndrome, a rare tumour predisposition syndrome. Cancer-associated hotspots mutations have been reported in both catalytic domains of Dicer, and are predicted to disrupt miRNA processing activity. To understand these hotspot mutations in cancer development, we have generated cell lines harbouring single amino acid substitutions within either the RNAse IIIa (S1344L) and the RNAse IIIb (D1709N) domains of the endogenous Dicer1 gene. We show that both mutations result in a widespread loss of 5p miRNAs, but, unexpectedly, an increase in 3p passenger strands loading into Ago2. Similarities between both mutants can be explained as the S1344 residue is structurally also part of the RNase IIIb catalytical site. Functionally, we found that changes in the repertoire of miRNAs loaded into Ago2 result in altered gene expression, impacting critical pathways for cancer development, including metastatic potential. Our results indicate that inactivating the processing activity of Dicer does not result in genomic instability. Instead, mutations cause a specific upregulation of human endogenous retrovirus H (HERVH) and 3p miRNAs, which have the potential to be used as markers for Dicer1 syndrome tumours.
2026-06-23 | PXD072167 | Pride
Project description:The DICER1 gene is mutated in cancer, including in DICER1-related tumour predisposition. Cancer-associated hotspot mutations have been reported in both catalytic domains of DICER and are predicted to disrupt miRNA biogenesis. To understand how these hotspot mutations contribute to cancer development, we generate cell lines harbouring single amino acid substitutions within the catalytic RNase IIIa (S1344L) or RNase IIIb (D1709N) domains of the endogenous DICER1 gene. Here we show that both mutations result in a widespread loss of 5p miRNAs, and an increase in 3p passenger strands loading into AGO2. The shared similarities between both mutants can be attributed to the structural proximity of the S1344 residue to the RNase IIIb catalytic centre. Functionally, we find that changes in the repertoire of miRNAs loaded into AGO2 result in altered gene expression, impacting critical pathways for cancer development, including metastatic potential. Additionally, our results indicate that inactivating the processing activity of DICER does not result in genomic instability. Instead, mutations cause upregulation of transposable elements, including the human endogenous retrovirus H through miRNA-independent mechanisms. This suggests that both canonical and non-canonical DICER functions are important to understand DICER1-related tumour predisposition.
2026-06-22 | GSE328604 | GEO
Project description:The DICER1 gene is mutated in cancer, including Dicer1 syndrome, a rare tumour predisposition syndrome. Cancer-associated hotspots mutations have been reported in both catalytic domains of Dicer, and are predicted to disrupt miRNA processing activity. To understand these hotspot mutations in cancer development, we have generated cell lines harbouring single amino acid substitutions within either the RNAse IIIa (S1344L) and the RNAse IIIb (D1709N) domains of the endogenous Dicer1 gene. We show that both mutations result in a widespread loss of 5p miRNAs, but, unexpectedly, an increase in 3p passenger strands loading into Ago2. Similarities between both mutants can be explained as the S1344 residue is structurally also part of the RNase IIIb catalytical site. Functionally, we found that changes in the repertoire of miRNAs loaded into Ago2 result in altered gene expression, impacting critical pathways for cancer development, including metastatic potential. Our results indicate that inactivating the processing activity of Dicer does not result in genomic instability. Instead, mutations cause a specific upregulation of human endogenous retrovirus H (HERVH) and 3p miRNAs, which have the potential to be used as markers for Dicer1 syndrome tumours.
2026-06-22 | GSE313187 | GEO
Project description:The DICER1 gene is mutated in cancer, including Dicer1 syndrome, a rare tumour predisposition syndrome. Cancer-associated hotspots mutations have been reported in both catalytic domains of Dicer, and are predicted to disrupt miRNA processing activity. To understand these hotspot mutations in cancer development, we have generated cell lines harbouring single amino acid substitutions within either the RNAse IIIa (S1344L) and the RNAse IIIb (D1709N) domains of the endogenous Dicer1 gene. We show that both mutations result in a widespread loss of 5p miRNAs, but, unexpectedly, an increase in 3p passenger strands loading into Ago2. Similarities between both mutants can be explained as the S1344 residue is structurally also part of the RNase IIIb catalytical site. Functionally, we found that changes in the repertoire of miRNAs loaded into Ago2 result in altered gene expression, impacting critical pathways for cancer development, including metastatic potential. Our results indicate that inactivating the processing activity of Dicer does not result in genomic instability. Instead, mutations cause a specific upregulation of human endogenous retrovirus H (HERVH) and 3p miRNAs, which have the potential to be used as markers for Dicer1 syndrome tumours.
2026-06-22 | GSE313185 | GEO
Project description:DICER1 plays a critical role in microRNA (miRNA) biogenesis. Recurrent somatic “hotspot” mutations at four mental binding sites within the RNase IIIb domain of DICER1, were identified in ovarian sex cord-stromal tumors and have since been described in other pediatric tumors. In this study, we identified and characterized DICER1 hotspot mutations in endometrial cancers derived from The Cancer Genome Atlas (TCGA) and our local tumor bank. DICER1 hotspot mutations are found in ~2% of endometrial tumors. Using Illumina and Sanger targeted resequencing we observed biallelic DICER1 mutations in more than 50% of cases with hotspot mutations and identified an additional recurrent mutation G1809R in 2 cases. Through small RNA deep sequencing and real-time PCR, we demonstrated mutations that add a positively charged side chain to residue 1809 have similar detrimental effects on 5p miRNA production as mutations at metal binding sites. In one case G1809R was compound heterozygous with a germline S839F mutation, which contributes to loss of DICER1 expression by promoting protein degradation. As expected, 5p miRNAs are globally reduced in tumors and cell lines with hotspot mutations. Pathway analysis of gene expression profiles indicated that genes derepressed due to loss of 5p miRNAs are strongly associated with cell cycle related pathways. Using a Dicer null cell line model, we demonstrated that DICER1 hotspot mutants abolished the inhibitory effects of wildtype DICER1 on cell proliferation upon re-expression. Furthermore, targets of let-7 family miRNAs are enriched among the upregulated genes, suggesting loss of let-7 may be impacting downstream pathways.
2015-03-16 | GSE65092 | GEO
Project description:The DICER1 gene is mutated in cancer, including Dicer1 syndrome, a rare tumour predisposition syndrome. Cancer-associated hotspots mutations have been reported in both catalytic domains of Dicer, and are predicted to disrupt miRNA processing activity. To understand these hotspot mutations in cancer development, we have generated cell lines harbouring single amino acid substitutions within either the RNAse IIIa (S1344L) and the RNAse IIIb (D1709N) domains of the endogenous Dicer1 gene. We show that both mutations result in a widespread loss of 5p miRNAs, but, unexpectedly, an increase in 3p passenger strands loading into Ago2. Similarities between both mutants can be explained as the S1344 residue is structurally also part of the RNase IIIb catalytical site. Functionally, we found that changes in the repertoire of miRNAs loaded into Ago2 result in altered gene expression, impacting critical pathways for cancer development, including metastatic potential. Our results indicate that inactivating the processing activity of Dicer does not result in genomic instability. Instead, mutations cause a specific upregulation of human endogenous retrovirus H (HERVH) and 3p miRNAs, which have the potential to be used as markers for Dicer1 syndrome tumours.
2026-06-22 | GSE313183 | GEO
Project description:Dicer is an essential enzyme in microRNA biogenesis. Mutations in the DICER1 gene are linked to various cancers, notably through the DICER1 syndrome. To investigate the impact of the pathogenic hotspot mutations in DICER1-associated tumors, we introduced a hotspot mutation into the endogenous Dicer1 locus of a mouse embryonic carcinoma cell line using CRISPR. Our findings not only confirm the loss of 5p-miRNAs, as previously reported, but also uncover an unexpected upregulation of specific 3p-miRNAs. These upregulated 3p-miRNAs, usually considered as passenger strands in the wild-type cells, are selectively loaded into the Argonaute protein in mutant cells based on their 5' end characteristics, resulting in a "strand-switch" phenomenon. Functional assays and transcriptome analyses demonstrate the passenger 3p-miRNAs’ activity. This study suggests that the Dicer hotspot mutation is not merely a loss-of-function mutation for 5p-miRNAs but also a gain-of-function mutation for passenger 3p-miRNA, potentially contributing to DICER1-associated tumorigenesis.
2025-07-18 | GSE279556 | GEO
Project description:Dicer is an essential enzyme in microRNA biogenesis. Mutations in the DICER1 gene are linked to various cancers, notably through the DICER1 syndrome. To investigate the impact of the pathogenic hotspot mutations in DICER1-associated tumors, we introduced a hotspot mutation into the endogenous Dicer1 locus of a mouse embryonic carcinoma cell line using CRISPR. Our findings not only confirm the loss of 5p-miRNAs, as previously reported, but also uncover an unexpected upregulation of specific 3p-miRNAs. These upregulated 3p-miRNAs, usually considered as passenger strands in the wild-type cells, are selectively loaded into the Argonaute protein in mutant cells based on their 5' end characteristics, resulting in a "strand-switch" phenomenon. Functional assays and transcriptome analyses demonstrate the passenger 3p-miRNAs’ activity. This study suggests that the Dicer hotspot mutation is not merely a loss-of-function mutation for 5p-miRNAs but also a gain-of-function mutation for passenger 3p-miRNA, potentially contributing to DICER1-associated tumorigenesis.
2025-07-18 | GSE279645 | GEO