Precise tuning of gene expression levels in mammalian cells.

Show simple item record Michaels, Yale S Barnkob, Mike B Barbosa, Hector Baeumler, Toni A Thompson, Mary K Andre, Violaine Colin-York, Huw Fritzsche, Marco Gileadi, Uzi Sheppard, Hilary M Knapp, David JHF Milne, Thomas A Cerundolo, Vincenzo Fulga, Tudor A
dc.coverage.spatial England 2022-06-21T02:25:19Z 2022-06-21T02:25:19Z 2019-02-18
dc.identifier.citation (2019). Nature Communications, 10(1), 818-.
dc.identifier.issn 2041-1723
dc.description.abstract Precise, analogue regulation of gene expression is critical for cellular function in mammals. In contrast, widely employed experimental and therapeutic approaches such as knock-in/out strategies are more suitable for binary control of gene activity. Here we report on a method for precise control of gene expression levels in mammalian cells using engineered microRNA response elements (MREs). First, we measure the efficacy of thousands of synthetic MRE variants under the control of an endogenous microRNA by high-throughput sequencing. Guided by this data, we establish a library of microRNA silencing-mediated fine-tuners (miSFITs) of varying strength that can be employed to precisely control the expression of user-specified genes. We apply this technology to tune the T-cell co-inhibitory receptor PD-1 and to explore how antigen expression influences T-cell activation and tumour growth. Finally, we employ CRISPR/Cas9 mediated homology directed repair to introduce miSFITs into the BRCA1 3'UTR, demonstrating that this versatile tool can be used to tune endogenous genes.
dc.format.medium Electronic
dc.language eng
dc.publisher Springer Science and Business Media LLC
dc.relation.ispartofseries Nature communications
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher.
dc.subject Animals
dc.subject Mice, Inbred C57BL
dc.subject Humans
dc.subject Melanoma, Experimental
dc.subject Ovalbumin
dc.subject Recombinant Proteins
dc.subject MicroRNAs
dc.subject 3' Untranslated Regions
dc.subject Xenograft Model Antitumor Assays
dc.subject Genetic Techniques
dc.subject Gene Expression Regulation
dc.subject Response Elements
dc.subject Genes, BRCA1
dc.subject HEK293 Cells
dc.subject High-Throughput Nucleotide Sequencing
dc.subject CRISPR-Cas Systems
dc.subject B7-H1 Antigen
dc.subject Genetics
dc.subject Biotechnology
dc.subject 1.1 Normal biological development and functioning
dc.subject 5.2 Cellular and gene therapies
dc.subject Cancer
dc.subject Science & Technology
dc.subject Multidisciplinary Sciences
dc.subject Science & Technology - Other Topics
dc.subject MICRORNA
dc.subject BINDING
dc.subject AFFINITY
dc.subject 0604 Genetics
dc.subject 0601 Biochemistry and Cell Biology
dc.subject Biomedical
dc.subject Basic Science
dc.title Precise tuning of gene expression levels in mammalian cells.
dc.type Journal Article
dc.identifier.doi 10.1038/s41467-019-08777-y
pubs.issue 1
pubs.begin-page 818
pubs.volume 10 2022-05-19T23:35:45Z
dc.rights.holder Copyright: The author en
dc.identifier.pmid 30778069 (pubmed)
pubs.publication-status Published
dc.rights.accessrights en
pubs.subtype Research Support, Non-U.S. Gov't
pubs.subtype research-article
pubs.subtype Journal Article
pubs.elements-id 764132 Science Biological Sciences Science Research Maurice Wilkins Centre (2010-2014)
dc.identifier.eissn 2041-1723
dc.identifier.pii 10.1038/s41467-019-08777-y
pubs.number 818
pubs.record-created-at-source-date 2022-05-20 2019-02-18

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