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A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells

Dgcr8 knockout cells provide a great means to understand the function of microRNAs (miRNAs) in vitro and in vivo. Current strategies to study miRNA function in Dgcr8 knockout cells depend on transient transfection of chemically synthesized miRNA mimics, which is costly and not suitable for long-term...

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Autores principales: Wang, Xi-Wen, Hao, Jing, Guo, Wen-Ting, Liao, Le-Qi, Huang, Si-Yue, Guo, Xiangpeng, Bao, Xichen, Esteban, Miguel A., Wang, Yangming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5830984/
https://www.ncbi.nlm.nih.gov/pubmed/28988987
http://dx.doi.org/10.1016/j.stemcr.2017.08.027
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author Wang, Xi-Wen
Hao, Jing
Guo, Wen-Ting
Liao, Le-Qi
Huang, Si-Yue
Guo, Xiangpeng
Bao, Xichen
Esteban, Miguel A.
Wang, Yangming
author_facet Wang, Xi-Wen
Hao, Jing
Guo, Wen-Ting
Liao, Le-Qi
Huang, Si-Yue
Guo, Xiangpeng
Bao, Xichen
Esteban, Miguel A.
Wang, Yangming
author_sort Wang, Xi-Wen
collection PubMed
description Dgcr8 knockout cells provide a great means to understand the function of microRNAs (miRNAs) in vitro and in vivo. Current strategies to study miRNA function in Dgcr8 knockout cells depend on transient transfection of chemically synthesized miRNA mimics, which is costly and not suitable for long-term study and genetic selection of miRNA function. Here, we developed a cost-effective DGCR8-independent stable miRNA expression (DISME) strategy based on a short hairpin RNA vector that can be precisely processed by DICER. Using DISME, we found that miR-294 promoted the formation of meso-endoderm lineages during embryonic stem cell differentiation. Furthermore, DISME allowed for a pooled screen of miRNA function and identified an miR-183–182 cluster of miRNAs promoting self-renewal and pluripotency in mouse embryonic stem cells. Altogether, our study demonstrates that DISME is a robust and cost-effective strategy that allows for long-term study and genetic selection of miRNA function in a Dgcr8 knockout background.
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spelling pubmed-58309842018-03-06 A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells Wang, Xi-Wen Hao, Jing Guo, Wen-Ting Liao, Le-Qi Huang, Si-Yue Guo, Xiangpeng Bao, Xichen Esteban, Miguel A. Wang, Yangming Stem Cell Reports Article Dgcr8 knockout cells provide a great means to understand the function of microRNAs (miRNAs) in vitro and in vivo. Current strategies to study miRNA function in Dgcr8 knockout cells depend on transient transfection of chemically synthesized miRNA mimics, which is costly and not suitable for long-term study and genetic selection of miRNA function. Here, we developed a cost-effective DGCR8-independent stable miRNA expression (DISME) strategy based on a short hairpin RNA vector that can be precisely processed by DICER. Using DISME, we found that miR-294 promoted the formation of meso-endoderm lineages during embryonic stem cell differentiation. Furthermore, DISME allowed for a pooled screen of miRNA function and identified an miR-183–182 cluster of miRNAs promoting self-renewal and pluripotency in mouse embryonic stem cells. Altogether, our study demonstrates that DISME is a robust and cost-effective strategy that allows for long-term study and genetic selection of miRNA function in a Dgcr8 knockout background. Elsevier 2017-10-05 /pmc/articles/PMC5830984/ /pubmed/28988987 http://dx.doi.org/10.1016/j.stemcr.2017.08.027 Text en © 2017 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Wang, Xi-Wen
Hao, Jing
Guo, Wen-Ting
Liao, Le-Qi
Huang, Si-Yue
Guo, Xiangpeng
Bao, Xichen
Esteban, Miguel A.
Wang, Yangming
A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title_full A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title_fullStr A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title_full_unstemmed A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title_short A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183–182 Families in Mouse Embryonic Stem Cells
title_sort dgcr8-independent stable microrna expression strategy reveals important functions of mir-290 and mir-183–182 families in mouse embryonic stem cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5830984/
https://www.ncbi.nlm.nih.gov/pubmed/28988987
http://dx.doi.org/10.1016/j.stemcr.2017.08.027
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