“Click handle”-modified 2′-deoxy-2′-fluoroarabino nucleic acid as a synthetic genetic polymer capable of post-polymerization functionalization

Abstract

The functions of natural nucleic acids such as DNA and RNA have transcended genetic information carriers and now encompass affinity reagents, molecular catalysts, nanostructures, data storage, and many others. However,...

From polymerase engineering to semi-synthetic life: artificial expansion of the central dogma

“…In another example, T7 RNAP mutant Y639F was shown to be efficient for the transcription of RNA with 2'-deoxy-2'-fluoro (5-ethynyl) uridine triphosphate and other natural NTPs, and used for the evolution of 2'-F-modified RNA-scaffolded carbohydrate clusters 67,168 . Very recently, Niu and co-workers synthesized C8-alkyne-FANA UTP, and demonstrated its enzymatic incorporation into FANA by Tgo DNAP 181 .This work further enriched the XNA toolbox with components containing clickable handles.…”

“…67,168 Very recently, Niu and co-workers synthesized C8-alkyne-FANA UTP, and demonstrated its enzymatic incorporation into FANA by Tgo DNAP. 181 This work further enriched the XNA toolbox with components containing clickable handles.…”

Advancing the enzymatic toolkit for 2′-fluoro arabino nucleic acid (FANA) manipulation: phosphorylation, ligation, replication, and templating RNA transcription

“…This integration offers a practical means to simultaneously leverage expanded functionality and the advantages of an XNA backbone. 22 To investigate the possibility of constructing DNA-FANA chimeric sequences via exponential amplication, we rst compared the rate of DNA-templated single nucleotide incorporation of fNTPs and dNTPs using Tgo polymerase decient in 3 0 -5 0 exonuclease activity [Tgo (exo-)]. When nucleotide concentrations were set at 300 and 600 nM, spanning routine dNTP concentrations in PCR, Tgo (exo-) displayed an incorporation rate for fATP and fGTP about 1.1-2 fold that of their DNA counterparts, similar incorporation rates for fCTP and dCTP, and a 2 to 4-fold reduction in the incorporation rates of fTTP and fUTP relative to their respective DNA counterparts (Fig.…”

Increasing the functional density of threose nucleic acid

“…Despite this potential weakness, click chemistry has been used to modify genetic polymers with a range of functional groups, including amino acids, carbohydrates, and dyes, and subsequent in vitro selection experiments have yielded aptamers against targets that were previously refractory to in vitro selection. 12–16…”

“…Despite this potential weakness, click chemistry has been used to modify genetic polymers with a range of functional groups, including amino acids, carbohydrates, and dyes, and subsequent in vitro selection experiments have yielded aptamers against targets that were previously refractory to in vitro selection. [12][13][14][15][16] The preparation of uniformly modified libraries requires synthetic strategies that introduce new chemical functionality directly into the oligonucleotide sequence during the library generation step of the selection. One approach involves the concept of genetic alphabet expansion, which increases the sequence diversity of the starting library through the incorporation of a third Watson-Crick base pair.…”