Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene
Maturation of microRNAs (miRNAs) begins by the “Microprocessor” complex, containing the Drosha endonuclease and its partner protein, "DiGeorge Syndrome Critical Region 8" (DGCR8). Although the main function of the two proteins is to coordinate the first step of precursor miRNAs formation,...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/13/11/1925 |
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| author | Dóra Reé Ábel Fóthi Nóra Varga Orsolya Kolacsek Tamás I. Orbán Ágota Apáti |
| author_facet | Dóra Reé Ábel Fóthi Nóra Varga Orsolya Kolacsek Tamás I. Orbán Ágota Apáti |
| author_sort | Dóra Reé |
| collection | DOAJ |
| description | Maturation of microRNAs (miRNAs) begins by the “Microprocessor” complex, containing the Drosha endonuclease and its partner protein, "DiGeorge Syndrome Critical Region 8" (DGCR8). Although the main function of the two proteins is to coordinate the first step of precursor miRNAs formation, several studies revealed their miRNA-independent functions in other RNA-related pathways (e.g., in snoRNA decay) or, for the DGCR8, the role in tissue development. To investigate the specific roles of DGCR8 in various cellular pathways, we previously established a human embryonic stem-cell (hESC) line carrying a monoallelic DGCR8 mutation by using the CRISPR-Cas9 system. In this study, we genetically characterized single-cell originated progenies of the cell line and showed that DGCR8 heterozygous mutation results in only a modest effect on the mRNA level but a significant decrease at the protein level. Self-renewal and trilineage differentiation capacity of these hESCs were not affected by the mutation. However, partial disturbance of the Microprocessor function could be revealed in pri-miRNA processing along the human chromosome 19 miRNA cluster in several clones. With all these studies, we can demonstrate that the mutant hESC line is a good model to study not only miRNA-related but also other “noncanonical” functions of the DGCR8 protein. |
| first_indexed | 2024-03-09T19:03:49Z |
| format | Article |
| id | doaj.art-b6250f8448ab4eebb4227542d2c016d9 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T19:03:49Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-b6250f8448ab4eebb4227542d2c016d92023-11-24T04:47:00ZengMDPI AGGenes2073-44252022-10-011311192510.3390/genes13111925Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 GeneDóra Reé0Ábel Fóthi1Nóra Varga2Orsolya Kolacsek3Tamás I. Orbán4Ágota Apáti5Institute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryInstitute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryInstitute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryInstitute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryInstitute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryInstitute of Enzymology, Research Center for Natural Sciences, 1117 Budapest, HungaryMaturation of microRNAs (miRNAs) begins by the “Microprocessor” complex, containing the Drosha endonuclease and its partner protein, "DiGeorge Syndrome Critical Region 8" (DGCR8). Although the main function of the two proteins is to coordinate the first step of precursor miRNAs formation, several studies revealed their miRNA-independent functions in other RNA-related pathways (e.g., in snoRNA decay) or, for the DGCR8, the role in tissue development. To investigate the specific roles of DGCR8 in various cellular pathways, we previously established a human embryonic stem-cell (hESC) line carrying a monoallelic DGCR8 mutation by using the CRISPR-Cas9 system. In this study, we genetically characterized single-cell originated progenies of the cell line and showed that DGCR8 heterozygous mutation results in only a modest effect on the mRNA level but a significant decrease at the protein level. Self-renewal and trilineage differentiation capacity of these hESCs were not affected by the mutation. However, partial disturbance of the Microprocessor function could be revealed in pri-miRNA processing along the human chromosome 19 miRNA cluster in several clones. With all these studies, we can demonstrate that the mutant hESC line is a good model to study not only miRNA-related but also other “noncanonical” functions of the DGCR8 protein.https://www.mdpi.com/2073-4425/13/11/1925miRNA processingC19MCmiRNA clusterhuman pluripotent stem cellsdifferentiation |
| spellingShingle | Dóra Reé Ábel Fóthi Nóra Varga Orsolya Kolacsek Tamás I. Orbán Ágota Apáti Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene Genes miRNA processing C19MC miRNA cluster human pluripotent stem cells differentiation |
| title | Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene |
| title_full | Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene |
| title_fullStr | Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene |
| title_full_unstemmed | Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene |
| title_short | Partial Disturbance of Microprocessor Function in Human Stem Cells Carrying a Heterozygous Mutation in the DGCR8 Gene |
| title_sort | partial disturbance of microprocessor function in human stem cells carrying a heterozygous mutation in the dgcr8 gene |
| topic | miRNA processing C19MC miRNA cluster human pluripotent stem cells differentiation |
| url | https://www.mdpi.com/2073-4425/13/11/1925 |
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