Modulating myoblast differentiation with RNA-based controllers
Tunable genetic controllers play a critical role in the engineering of biological systems that respond to environmental and cellular signals. RNA devices, a class of engineered RNA-based controllers, enable tunable gene expression control of target genes in response to molecular effectors. RNA devic...
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| Veröffentlicht in: | PloS one 2022-09, Vol.17 (9), p.e0275298 |
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| creator | Dykstra, Peter B Rando, Thomas A Smolke, Christina D |
| description | Tunable genetic controllers play a critical role in the engineering of biological systems that respond to environmental and cellular signals. RNA devices, a class of engineered RNA-based controllers, enable tunable gene expression control of target genes in response to molecular effectors. RNA devices have been demonstrated in a number of systems showing proof-of-concept of applying ligand-responsive control over therapeutic activities, including regulation of cell fate decisions such as T cell proliferation and apoptosis. Here, we describe the application of a theophylline-responsive RNA device in a muscle progenitor cell system to control myogenic differentiation. Ribozyme-based RNA switches responsive to theophylline control fluorescent reporter expression in C2C12 myoblasts in a ligand dependent manner. HRAS and JAK1, both anti-differentiation proteins, were incorporated into RNA devices. Finally, we demonstrate that the regulation of HRAS expression via theophylline-responsive RNA devices results in the modulation of myoblast differentiation in a theophylline-dependent manner. Our work highlights the potential for RNA devices to exert drug-responsive, tunable control over cell fate decisions with applications in stem cell therapy and basic stem cell biology research. |
| doi_str_mv | 10.1371/journal.pone.0275298 |
| format | Article |
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RNA devices, a class of engineered RNA-based controllers, enable tunable gene expression control of target genes in response to molecular effectors. RNA devices have been demonstrated in a number of systems showing proof-of-concept of applying ligand-responsive control over therapeutic activities, including regulation of cell fate decisions such as T cell proliferation and apoptosis. Here, we describe the application of a theophylline-responsive RNA device in a muscle progenitor cell system to control myogenic differentiation. Ribozyme-based RNA switches responsive to theophylline control fluorescent reporter expression in C2C12 myoblasts in a ligand dependent manner. HRAS and JAK1, both anti-differentiation proteins, were incorporated into RNA devices. Finally, we demonstrate that the regulation of HRAS expression via theophylline-responsive RNA devices results in the modulation of myoblast differentiation in a theophylline-dependent manner. Our work highlights the potential for RNA devices to exert drug-responsive, tunable control over cell fate decisions with applications in stem cell therapy and basic stem cell biology research.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0275298</identifier><identifier>PMID: 36166456</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Apoptosis ; Biology and Life Sciences ; Catalytic RNA ; Cell differentiation ; Cell Differentiation - genetics ; Cell fate ; Cell proliferation ; Cell therapy ; Controllers ; Decisions ; Design ; Devices ; Differentiation ; Dosage and administration ; Fluorescence ; Gene expression ; Genetic engineering ; Health aspects ; Janus kinase ; Ligands ; Lymphocytes ; Lymphocytes T ; Methods ; Muscle Development - genetics ; Muscles ; Musculoskeletal system ; Myoblasts ; Myoblasts - metabolism ; Progenitor cells ; Protein expression ; Proteins ; Research and Analysis Methods ; Ribonucleic acid ; RNA ; RNA - genetics ; RNA - metabolism ; RNA, Catalytic - metabolism ; Sensors ; Stem cells ; Switches ; Synthetic biology ; Theophylline ; Theophylline - metabolism ; Theophylline - pharmacology ; Tissue engineering ; Transplantation ; Transplants & implants</subject><ispartof>PloS one, 2022-09, Vol.17 (9), p.e0275298</ispartof><rights>COPYRIGHT 2022 Public Library of Science</rights><rights>2022 Dykstra et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2022 Dykstra et al 2022 Dykstra et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-e96b92a3d98ea8e542acc05729a41892e355d8a3ad8a93d5a0b064274952b4503</citedby><cites>FETCH-LOGICAL-c692t-e96b92a3d98ea8e542acc05729a41892e355d8a3ad8a93d5a0b064274952b4503</cites><orcidid>0000-0002-5449-8495 ; 0000-0002-2626-7104</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514614/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514614/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36166456$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Preiss, Thomas</contributor><creatorcontrib>Dykstra, Peter B</creatorcontrib><creatorcontrib>Rando, Thomas A</creatorcontrib><creatorcontrib>Smolke, Christina D</creatorcontrib><title>Modulating myoblast differentiation with RNA-based controllers</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Tunable genetic controllers play a critical role in the engineering of biological systems that respond to environmental and cellular signals. RNA devices, a class of engineered RNA-based controllers, enable tunable gene expression control of target genes in response to molecular effectors. RNA devices have been demonstrated in a number of systems showing proof-of-concept of applying ligand-responsive control over therapeutic activities, including regulation of cell fate decisions such as T cell proliferation and apoptosis. Here, we describe the application of a theophylline-responsive RNA device in a muscle progenitor cell system to control myogenic differentiation. Ribozyme-based RNA switches responsive to theophylline control fluorescent reporter expression in C2C12 myoblasts in a ligand dependent manner. HRAS and JAK1, both anti-differentiation proteins, were incorporated into RNA devices. Finally, we demonstrate that the regulation of HRAS expression via theophylline-responsive RNA devices results in the modulation of myoblast differentiation in a theophylline-dependent manner. 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Our work highlights the potential for RNA devices to exert drug-responsive, tunable control over cell fate decisions with applications in stem cell therapy and basic stem cell biology research.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>36166456</pmid><doi>10.1371/journal.pone.0275298</doi><tpages>e0275298</tpages><orcidid>https://orcid.org/0000-0002-5449-8495</orcidid><orcidid>https://orcid.org/0000-0002-2626-7104</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Analysis Apoptosis Biology and Life Sciences Catalytic RNA Cell differentiation Cell Differentiation - genetics Cell fate Cell proliferation Cell therapy Controllers Decisions Design Devices Differentiation Dosage and administration Fluorescence Gene expression Genetic engineering Health aspects Janus kinase Ligands Lymphocytes Lymphocytes T Methods Muscle Development - genetics Muscles Musculoskeletal system Myoblasts Myoblasts - metabolism Progenitor cells Protein expression Proteins Research and Analysis Methods Ribonucleic acid RNA RNA - genetics RNA - metabolism RNA, Catalytic - metabolism Sensors Stem cells Switches Synthetic biology Theophylline Theophylline - metabolism Theophylline - pharmacology Tissue engineering Transplantation Transplants & implants |
| title | Modulating myoblast differentiation with RNA-based controllers |
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