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Exploring of the feature space of de novo developed post-transcriptional riboregulators

Gert Peters (UGent) , Jo Maertens (UGent) , Jeroen Lammertyn and Marjan De Mey (UGent)
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Abstract
Metabolic engineering increasingly depends upon RNA technology to customly rewire the metabolism to maximize production. To this end, pure riboregulators allow dynamic gene repression without the need of a potentially burdensome coexpressed protein like typical Hfq binding small RNAs and clustered regularly interspaced short palindromic repeats technology. Despite this clear advantage, no clear general design principles are available to de novo develop repressing riboregulators, limiting the availability and the reliable development of these type of riboregulators. Here, to overcome this lack of knowledge on the functionality of repressing riboregulators, translation inhibiting RNAs are developed from scratch. These de novo developed riboregulators explore features related to thermodynamical and structural factors previously attributed to translation initiation modulation. In total, 12 structural and thermodynamic features were defined of which six features were retained after removing correlations from an in silico generated riboregulator library. From this translation inhibiting RNA library, 18 riboregulators were selected using a experimental design and subsequently constructed and co-expressed with two target untranslated regions to link the translation inhibiting RNA features to functionality. The pure riboregulators in the design of experiments showed repression down to 6% of the original protein expression levels, which could only be partially explained by a ordinary least squares regression model. To allow reliable forward engineering, a partial least squares regression model was constructed and validated to link the properties of translation inhibiting RNA riboregulators to gene repression. In this model both structural and thermodynamic features were important for efficient gene repression by pure riboregulators. This approach enables a more reliable de novo forward engineering of effective pure riboregulators, which further expands the RNA toolbox for gene expression modulation.
Keywords
TRANSCRIPTION ACTIVATING RNAS, ESCHERICHIA-COLI, GENE-EXPRESSION, PROTEIN EXPRESSION, QUANTITATIVE-ANALYSIS, SECONDARY STRUCTURES, AUTOMATED DESIGN, ANTISENSE RNAS, REGULATORS, PATHWAY

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MLA
Peters, Gert, Jo Maertens, Jeroen Lammertyn, et al. “Exploring of the Feature Space of De Novo Developed Post-transcriptional Riboregulators.” Ed. Shi-Jie Chen. PLOS COMPUTATIONAL BIOLOGY 14.8 (2018): n. pag. Print.
APA
Peters, G., Maertens, J., Lammertyn, J., & De Mey, M. (2018). Exploring of the feature space of de novo developed post-transcriptional riboregulators. (S.-J. Chen, Ed.)PLOS COMPUTATIONAL BIOLOGY, 14(8).
Chicago author-date
Peters, Gert, Jo Maertens, Jeroen Lammertyn, and Marjan De Mey. 2018. “Exploring of the Feature Space of De Novo Developed Post-transcriptional Riboregulators.” Ed. Shi-Jie Chen. Plos Computational Biology 14 (8).
Chicago author-date (all authors)
Peters, Gert, Jo Maertens, Jeroen Lammertyn, and Marjan De Mey. 2018. “Exploring of the Feature Space of De Novo Developed Post-transcriptional Riboregulators.” Ed. Shi-Jie Chen. Plos Computational Biology 14 (8).
Vancouver
1.
Peters G, Maertens J, Lammertyn J, De Mey M. Exploring of the feature space of de novo developed post-transcriptional riboregulators. Chen S-J, editor. PLOS COMPUTATIONAL BIOLOGY. 2018;14(8).
IEEE
[1]
G. Peters, J. Maertens, J. Lammertyn, and M. De Mey, “Exploring of the feature space of de novo developed post-transcriptional riboregulators,” PLOS COMPUTATIONAL BIOLOGY, vol. 14, no. 8, 2018.
@article{8571262,
  abstract     = {Metabolic engineering increasingly depends upon RNA technology to customly rewire the metabolism to maximize production. To this end, pure riboregulators allow dynamic gene repression without the need of a potentially burdensome coexpressed protein like typical Hfq binding small RNAs and clustered regularly interspaced short palindromic repeats technology. Despite this clear advantage, no clear general design principles are available to de novo develop repressing riboregulators, limiting the availability and the reliable development of these type of riboregulators. Here, to overcome this lack of knowledge on the functionality of repressing riboregulators, translation inhibiting RNAs are developed from scratch. These de novo developed riboregulators explore features related to thermodynamical and structural factors previously attributed to translation initiation modulation. In total, 12 structural and thermodynamic features were defined of which six features were retained after removing correlations from an in silico generated riboregulator library. From this translation inhibiting RNA library, 18 riboregulators were selected using a experimental design and subsequently constructed and co-expressed with two target untranslated regions to link the translation inhibiting RNA features to functionality. The pure riboregulators in the design of experiments showed repression down to 6% of the original protein expression levels, which could only be partially explained by a ordinary least squares regression model. To allow reliable forward engineering, a partial least squares regression model was constructed and validated to link the properties of translation inhibiting RNA riboregulators to gene repression. In this model both structural and thermodynamic features were important for efficient gene repression by pure riboregulators. This approach enables a more reliable de novo forward engineering of effective pure riboregulators, which further expands the RNA toolbox for gene expression modulation.},
  articleno    = {e1006170},
  author       = {Peters, Gert and Maertens, Jo and Lammertyn, Jeroen and De Mey, Marjan},
  editor       = {Chen, Shi-Jie},
  issn         = {1553-7358},
  journal      = {PLOS COMPUTATIONAL BIOLOGY},
  keywords     = {TRANSCRIPTION ACTIVATING RNAS,ESCHERICHIA-COLI,GENE-EXPRESSION,PROTEIN EXPRESSION,QUANTITATIVE-ANALYSIS,SECONDARY STRUCTURES,AUTOMATED DESIGN,ANTISENSE RNAS,REGULATORS,PATHWAY},
  language     = {eng},
  number       = {8},
  pages        = {19},
  title        = {Exploring of the feature space of de novo developed post-transcriptional riboregulators},
  url          = {http://dx.doi.org/10.1371/journal.pcbi.1006170},
  volume       = {14},
  year         = {2018},
}

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