In situ probe and inhibitory RNA synthesis using streamlined gene cloning with Gibson assembly

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dc.contributor.authorWolff, Andrew
dc.contributor.authorWagner, Cynthia
dc.contributor.authorWolf, Julia
dc.contributor.authorLobo, Daniel
dc.date.accessioned2024-03-06T18:52:26Z
dc.date.available2024-03-06T18:52:26Z
dc.date.issued2022-06-14
dc.description.abstractThe synthesis of single-stranded riboprobes or double-stranded RNAs for in situ hybridization and gene knockdowns often use vectors that require time-consuming plasmid restriction digests and inefficient gel purifications. Here, we present a faster protocol for the simultaneous plasmid restriction digestion and Gibson assembly of vectors for the synthesis of both riboprobes and double-stranded RNAs for in situ and RNA interference experiments, respectively. We illustrate the protocol with planaria in situ and RNAi assays, but it is applicable to any organism.
dc.description.sponsorshipWe thank Erin Davies, Tim Demtro¨ der, Tagide deCarvalho, and the members of the Lobo Lab for helpful discussions. This work was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number R35GM137953. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
dc.description.urihttps://www.sciencedirect.com/science/article/pii/S2666166722003380
dc.format.extent17 pages
dc.genrejournal articles
dc.identifierdoi:10.13016/m2un0h-wxcs
dc.identifier.citationWolff, Andrew, Cynthia Wagner, Julia Wolf, and Daniel Lobo. “In Situ Probe and Inhibitory RNA Synthesis Using Streamlined Gene Cloning with Gibson Assembly.” STAR Protocols 3, no. 3 (September 16, 2022): 101458. https://doi.org/10.1016/j.xpro.2022.101458.
dc.identifier.urihttps://doi.org/10.1016/j.xpro.2022.101458
dc.identifier.urihttp://hdl.handle.net/11603/31859
dc.language.isoen_US
dc.publisherCell Press
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Faculty Collection
dc.relation.ispartofUMBC Biological Sciences Department
dc.rightsThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.
dc.rightsCreative Commons Attribution 4.0 International (CC BY 4.0)
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectDevelopmental biology
dc.subjectModel Organisms
dc.subjectMolecular Biology
dc.titleIn situ probe and inhibitory RNA synthesis using streamlined gene cloning with Gibson assembly
dc.typeText
dcterms.creatorhttps://orcid.org/0000-0002-9677-219X
dcterms.creatorhttps://orcid.org/0000-0003-4103-6926
dcterms.creatorhttps://orcid.org/0000-0003-4666-6118

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