Engineering programmable trans-splicing riboregulators for complex cellular logic computation
dc.contributor.advisor
French, Chris
dc.contributor.advisor
Granneman, Sander
dc.contributor.author
Gao, Yuanli
dc.contributor.sponsor
Darwin Trust of Edinburgh
en
dc.date.accessioned
2025-01-15T14:13:57Z
dc.date.available
2025-01-15T14:13:57Z
dc.date.issued
2025-01-15
dc.description.abstract
Synthetic genetic circuits program the cellular input-output relationships to execute customized functions. However, efforts to scale up these circuits have been hampered by the limited number of reliable regulatory mechanisms with high programmability, performance, predictability, and orthogonality. In this thesis, I engineered a class of Split-intron ENabled Trans-splicing Riboregulators (SENTRs) based on de-novo-designed external guide sequences (EGSs). SENTR libraries provide low leakage expression, wide dynamic range, and low crosstalk at multiple component levels. SENTRs can sense RNA targets, process signals by logic computation, and transduce them into various outputs, either mRNAs or ncRNAs. SENTR represents a powerful and versatile regulatory tool at the post-transcriptional level in Escherichia coli, with broad applications in biotechnology. I subsequently demonstrated that digital logic operation with up to six inputs could be implemented by using multiple orthogonal SENTRs to regulate a single gene simultaneously and coupling SENTRs with split intein-mediated protein trans-splicing. The integration of split introns and inteins is an efficient, genetically compact, and generally applicable strategy for building multi-input processing devices to significantly increase the information processing capacity of a single regulator gene.
en
dc.identifier.uri
https://hdl.handle.net/1842/43001
dc.identifier.uri
http://dx.doi.org/10.7488/era/5552
dc.language.iso
en
en
dc.publisher
The University of Edinburgh
en
dc.relation.hasversion
Gao, Y., Wang, L. & Wang, B. Customizing cellular signal processing by synthetic multi-level regulatory circuits. Nat. Commun. 14, 8415 (2023)
en
dc.relation.hasversion
Gao, X. J., Chong, L. S., Kim, M. S. & Elowitz, M. B. Programmable protein circuits in living cells. Science 361, 1252–1258 (2018)
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dc.subject
Synthetic Biology
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dc.subject
Genetic circuit
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dc.subject
Intron
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dc.subject
Biocomputation
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dc.subject
RNA splicing
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dc.title
Engineering programmable trans-splicing riboregulators for complex cellular logic computation
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dc.type
Thesis or Dissertation
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dc.type.qualificationlevel
Doctoral
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dc.type.qualificationname
PhD Doctor of Philosophy
en
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