Silver: Assembly: Difference between revisions
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Unique nucleotide sequence (UNS)-guided DNA assembly is a variation on Gibson/isothermal assembly that is more tolerant to repeated sequence elements, e.g. those common in synthetic metabolic pathways and genetic circuits. In this approach, 40 bp UNSes are algorithmically designed to be biologically inert and to efficiently undergo isothermal assembly, then attached to the termini of the DNA parts to be assembled, thereby providing unique and efficient homology regions for assembly. | Unique nucleotide sequence (UNS)-guided DNA assembly is a variation on Gibson/isothermal assembly that is more tolerant to repeated sequence elements, e.g. those common in synthetic metabolic pathways and genetic circuits. In this approach, 40 bp UNSes are algorithmically designed to be biologically inert and to efficiently undergo isothermal assembly, then attached to the termini of the DNA parts to be assembled, thereby providing unique and efficient homology regions for assembly. | ||
We first elaborated the details of this approach, and demonstrated its utility for metabolic engineering, in [http://nar.oxfordjournals.org/content/42/1/681.long Torella et al. 2014, "Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly."] We also used this approach to build mammalian genetic circuits in [http://nar.oxfordjournals.org/content/41/21/9967.long Lienert et al. 2014, "Two- and three-input TALE-based AND logic computation in embryonic stem cells."] | We first elaborated the details of this approach, and demonstrated its utility for metabolic engineering, in [http://nar.oxfordjournals.org/content/42/1/681.long Torella et al. 2014, "Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly."] | ||
We have also used this approach to build mammalian genetic circuits in [http://nar.oxfordjournals.org/content/41/21/9967.long Lienert et al. 2014, "Two- and three-input TALE-based AND logic computation in embryonic stem cells."] | |||
Revision as of 12:19, 14 April 2014
Unique nucleotide sequence (UNS)-guided DNA assembly is a variation on Gibson/isothermal assembly that is more tolerant to repeated sequence elements, e.g. those common in synthetic metabolic pathways and genetic circuits. In this approach, 40 bp UNSes are algorithmically designed to be biologically inert and to efficiently undergo isothermal assembly, then attached to the termini of the DNA parts to be assembled, thereby providing unique and efficient homology regions for assembly.
We first elaborated the details of this approach, and demonstrated its utility for metabolic engineering, in Torella et al. 2014, "Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly."
We have also used this approach to build mammalian genetic circuits in Lienert et al. 2014, "Two- and three-input TALE-based AND logic computation in embryonic stem cells."
