Difference between revisions of "Part:BBa K3154002"
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| − | + | Toehold switches are a class of de novo designed mRNA based riboregulators that possess unique properties such as low crosstalk, high orthogonality and high dynamic range. They are hence highly desirable as agents of translational regulation of reporter protein genes in abiotic situations for many synthetic biology applications. Toehold switches have free energy and sequence constraints amongst the various domains that determine their optimal conformation. The second generation toehold switch design allows for higher programmability of the toehold switch sequences, resulting in better performance. | |
| + | The trigger sequence for second generation designs are comprised of two sequences, resulting in a hybrid structure. An antisense RNA, or antimiR, is designed such that it is partially complementary to the microRNA. This creates a hybrid, T shaped trigger sequence where the base paired stem of 12 nt length sequesters the stop codon, and the free ends of 22 nt in total, form a complex with the toehold switch. The 5’ end of the toehold switch, or the unpaired toehold domain is complementary to the 3’ segment of the hybrid trigger consisting of the miR and the 5’ end of the hybrid trigger is the customized antimiR complex that is complementary to the bottom stem sequence. The unfolding of the toehold switch into a linear mRNA molecule is conditional on the base pairing action of the hybrid trigger with the switch region. Thus, second generation toehold switch design has an essential step prior to switch - trigger complex formation: the miR-antimiR complex formation. | ||
Revision as of 21:41, 15 October 2019
Toehold switches are a class of de novo designed mRNA based riboregulators that possess unique properties such as low crosstalk, high orthogonality and high dynamic range. They are hence highly desirable as agents of translational regulation of reporter protein genes in abiotic situations for many synthetic biology applications. Toehold switches have free energy and sequence constraints amongst the various domains that determine their optimal conformation. The second generation toehold switch design allows for higher programmability of the toehold switch sequences, resulting in better performance. The trigger sequence for second generation designs are comprised of two sequences, resulting in a hybrid structure. An antisense RNA, or antimiR, is designed such that it is partially complementary to the microRNA. This creates a hybrid, T shaped trigger sequence where the base paired stem of 12 nt length sequesters the stop codon, and the free ends of 22 nt in total, form a complex with the toehold switch. The 5’ end of the toehold switch, or the unpaired toehold domain is complementary to the 3’ segment of the hybrid trigger consisting of the miR and the 5’ end of the hybrid trigger is the customized antimiR complex that is complementary to the bottom stem sequence. The unfolding of the toehold switch into a linear mRNA molecule is conditional on the base pairing action of the hybrid trigger with the switch region. Thus, second generation toehold switch design has an essential step prior to switch - trigger complex formation: the miR-antimiR complex formation.