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Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN

MicroRNAs are important gene regulators involved in many biological processes, including stemness maintenance and cellular reprogramming. Current methods used in loss-of-function studies of microRNAs mainly include locked nucleic acid (LNA) oligonucleotides and miRZip inhibitors, which have several...

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Autores principales: Zhang, Zhonghui, Xiang, Di, Heriyanto, Fransisca, Gao, Yongxing, Qian, Zhijian, Wu, Wen-Shu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3849252/
https://www.ncbi.nlm.nih.gov/pubmed/24319658
http://dx.doi.org/10.1016/j.stemcr.2013.07.002
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author Zhang, Zhonghui
Xiang, Di
Heriyanto, Fransisca
Gao, Yongxing
Qian, Zhijian
Wu, Wen-Shu
author_facet Zhang, Zhonghui
Xiang, Di
Heriyanto, Fransisca
Gao, Yongxing
Qian, Zhijian
Wu, Wen-Shu
author_sort Zhang, Zhonghui
collection PubMed
description MicroRNAs are important gene regulators involved in many biological processes, including stemness maintenance and cellular reprogramming. Current methods used in loss-of-function studies of microRNAs mainly include locked nucleic acid (LNA) oligonucleotides and miRZip inhibitors, which have several limitations. Due to their unique gene structures and small sizes, there is no efficient or simple strategy to knock down or knock out microRNAs or whole microRNA clusters. Here, we demonstrate knockdown of the miR-302/367 cluster by using the Kruppel-associated box repressor domain fused with specific transcription activator-like effectors (TALEs) designed to bind the miR-302/367 cluster promoter. We also designed two pairs of TALE nucleases (TALENs) to efficiently delete the miR-302/367 cluster in primary human fibroblasts and determined that knockout of the miR-302/367 cluster completely blocked induced pluripotent stem cell (iPSC) generation. Together, our results demonstrate that TALE-based transcriptional repressor and TALENs are two promising approaches for loss-of-function studies of microRNA clusters in somatic cells and pluripotent stem cells.
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spelling pubmed-38492522013-12-06 Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN Zhang, Zhonghui Xiang, Di Heriyanto, Fransisca Gao, Yongxing Qian, Zhijian Wu, Wen-Shu Stem Cell Reports Report MicroRNAs are important gene regulators involved in many biological processes, including stemness maintenance and cellular reprogramming. Current methods used in loss-of-function studies of microRNAs mainly include locked nucleic acid (LNA) oligonucleotides and miRZip inhibitors, which have several limitations. Due to their unique gene structures and small sizes, there is no efficient or simple strategy to knock down or knock out microRNAs or whole microRNA clusters. Here, we demonstrate knockdown of the miR-302/367 cluster by using the Kruppel-associated box repressor domain fused with specific transcription activator-like effectors (TALEs) designed to bind the miR-302/367 cluster promoter. We also designed two pairs of TALE nucleases (TALENs) to efficiently delete the miR-302/367 cluster in primary human fibroblasts and determined that knockout of the miR-302/367 cluster completely blocked induced pluripotent stem cell (iPSC) generation. Together, our results demonstrate that TALE-based transcriptional repressor and TALENs are two promising approaches for loss-of-function studies of microRNA clusters in somatic cells and pluripotent stem cells. Elsevier 2013-08-15 /pmc/articles/PMC3849252/ /pubmed/24319658 http://dx.doi.org/10.1016/j.stemcr.2013.07.002 Text en © 2013 The Authors https://creativecommons.org/licenses/by-nc-nd/3.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License (https://creativecommons.org/licenses/by-nc-nd/3.0/) .
spellingShingle Report
Zhang, Zhonghui
Xiang, Di
Heriyanto, Fransisca
Gao, Yongxing
Qian, Zhijian
Wu, Wen-Shu
Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title_full Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title_fullStr Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title_full_unstemmed Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title_short Dissecting the Roles of miR-302/367 Cluster in Cellular Reprogramming Using TALE-based Repressor and TALEN
title_sort dissecting the roles of mir-302/367 cluster in cellular reprogramming using tale-based repressor and talen
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3849252/
https://www.ncbi.nlm.nih.gov/pubmed/24319658
http://dx.doi.org/10.1016/j.stemcr.2013.07.002
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