Efficiency of the synthetic self-splicing RiboJ ribozyme is robust to cis- and trans-changes in genetic background

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dc.citation.issue4
dc.citation.volume10
dc.contributor.authorVlková M
dc.contributor.authorMorampalli BR
dc.contributor.authorSilander OK
dc.coverage.spatialEngland
dc.date.accessioned2024-01-09T01:54:11Z
dc.date.accessioned2024-07-25T06:43:36Z
dc.date.available2021-03-18
dc.date.available2024-01-09T01:54:11Z
dc.date.available2024-07-25T06:43:36Z
dc.date.issued2021-08-24
dc.description.abstractThe expanding knowledge of the variety of synthetic genetic elements has enabled the construction of new and more efficient genetic circuits and yielded novel insights into molecular mechanisms. However, context dependence, in which interactions between cis- or trans-genetic elements affect the behavior of these elements, can reduce their general applicability or predictability. Genetic insulators, which mitigate unintended context-dependent cis-interactions, have been used to address this issue. One of the most commonly used genetic insulators is a self-splicing ribozyme called RiboJ, which can be used to decouple upstream 5' UTR in mRNA from downstream sequences (e.g., open reading frames). Despite its general use as an insulator, there has been no systematic study quantifying the efficiency of RiboJ splicing or whether this autocatalytic activity is robust to trans- and cis-genetic context. Here, we determine the robustness of RiboJ splicing in the genetic context of six widely divergent E. coli strains. We also check for possible cis-effects by assessing two SNP versions close to the catalytic site of RiboJ. We show that mRNA molecules containing RiboJ are rapidly spliced even during rapid exponential growth and high levels of gene expression, with a mean efficiency of 98%. We also show that neither the cis- nor trans-genetic context has a significant impact on RiboJ activity, suggesting this element is robust to both cis- and trans-genetic changes.
dc.description.confidentialfalse
dc.edition.editionAugust 2021
dc.format.paginatione1232-
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/34459545
dc.identifier.citationVlková M, Morampalli BR, Silander OK. (2021). Efficiency of the synthetic self-splicing RiboJ ribozyme is robust to cis- and trans-changes in genetic background.. Microbiologyopen. 10. 4. (pp. e1232-).
dc.identifier.doi10.1002/mbo3.1232
dc.identifier.eissn2045-8827
dc.identifier.elements-typejournal-article
dc.identifier.issn2045-8827
dc.identifier.numberARTN e1232
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/70753
dc.languageeng
dc.publisherJohn Wiley and Sons, Ltd
dc.publisher.urihttps://onlinelibrary.wiley.com/doi/10.1002/mbo3.1232
dc.relation.isPartOfMicrobiologyopen
dc.rights(c) The author/sen
dc.rights.licenseCC BYen
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectRT-qPCR
dc.subjectRiboJ
dc.subjectinsulation
dc.subjectribozyme
dc.subjectsplicing
dc.subject5' Untranslated Regions
dc.subjectEscherichia coli
dc.subjectGene Expression Regulation, Bacterial
dc.subjectGenome, Bacterial
dc.subjectLac Operon
dc.subjectOpen Reading Frames
dc.subjectPlasmids
dc.subjectPolymorphism, Single Nucleotide
dc.subjectPromoter Regions, Genetic
dc.subjectRNA Splicing
dc.subjectRNA, Catalytic
dc.subjectRNA, Messenger
dc.titleEfficiency of the synthetic self-splicing RiboJ ribozyme is robust to cis- and trans-changes in genetic background
dc.typeJournal article
pubs.elements-id448324
pubs.organisational-groupOther
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