Topics in medicinal chemistry, Год журнала: 2024, Номер unknown
Опубликована: Янв. 1, 2024
Язык: Английский
Chemical Society Reviews, Год журнала: 2025, Номер unknown
Опубликована: Янв. 1, 2025
The deep interconnection between nucleic acids and carbohydrates has prompted numerous synthetic campaigns to understand acid etiology, develop orthogonal genetic systems, improve the properties of therapeutic oligonucleotides.
Язык: Английский
Процитировано
2
Biotechnology Journal, Год журнала: 2024, Номер 19(9)
Опубликована: Сен. 1, 2024
Terminal deoxynucleotidyl transferase (TdT), a unique DNA polymerase that catalyzes the template-free incorporation of nucleotides into single-stranded DNA, has facilitated development various oligonucleotide-based tools and methods, especially in field enzymatic synthesis. However, expressing vertebrate-derived TdTs Escherichia coli complicates purification increases production costs. In this study, N-terminal truncation was performed to improve their expression stability. The results revealed could enhance level six TdTs. Among truncated mutants, N-140-ZaTdT N-140-CpTdT, with 140 amino acids removed, exhibited an increase protein expression, which 9.5- 23-fold higher than wild-types, respectively. Importantly, preserves catalytic function TdT. Additionally, T
Язык: Английский
Процитировано
3
Talanta, Год журнала: 2025, Номер 287, С. 127589 - 127589
Опубликована: Янв. 13, 2025
Язык: Английский
Процитировано
0
Biochemical Journal, Год журнала: 2025, Номер 482(2), С. 39 - 56
Опубликована: Янв. 1, 2025
DNA-joining by ligase and polymerase enzymes has provided the foundational tools for generating recombinant DNA enabled assembly of gene genome-sized synthetic products. Xenobiotic nucleic acid (XNA) analogues RNA with alternatives to canonical bases, so-called 'unnatural' nucleobase pairs (UBP-XNAs), represent next frontier technologies, applications as novel therapeutics in engineering semi-synthetic biological organisms. To realise full potential UBP-XNAs, researchers require a suite compatible processing acids on par those already available manipulating DNA. In particular, able join UBP-XNA will be essential large assemblies also hold promise synthesis single-stranded oligonucleotides. Here, we review recent emerging advances enzymes, polymerases ligases, describe their manipulation. We discuss future directions this field which consider involve two-pronged approaches enzyme biodiscovery natural coupled improvement structure-guided engineering.
Язык: Английский
Процитировано
0
ACS Synthetic Biology, Год журнала: 2025, Номер unknown
Опубликована: Янв. 29, 2025
Cell-free synthetic biology incorporates purified components and/or crude cell extracts to carry out metabolic and genetic programs. While protein synthesis has historically been the primary focus, more metabolism researchers are now turning toward cell-free systems either prototype pathways for cellular implementation or design new-to-nature reaction networks that incorporate environmentally relevant substrates new energy sources. The ability design, build, test enzyme combinations in vitro accelerated efforts understand bottlenecks engineer high-yielding pathways. However, only a small fraction of possibilities explored systems, from model organisms remain most common starting points. Expanding scope include organisms, alternative pathways, non-natural chemistries will enhance our bio-based chemical conversions.
Язык: Английский
Процитировано
0
Nucleic Acids Research, Год журнала: 2025, Номер 53(6)
Опубликована: Март 20, 2025
Abstract Given the emerging use of terminal deoxynucleotidyl transferase (TdT) in biotechnology and its clinical potential as a cancer marker target, development versatile probe system to study processivity, substrate properties, inhibition is highly desired. Here, we demonstrate multilayered application series environment-sensitive fluorescent 2′-deoxynucleotide probes that harness activity TdT accessing site-specifically functionalized DNA oligonucleotides devising real-time fluorescence platform monitor enzyme identify inhibitors. The nucleotides constructed by coupling heterocycles progressively increasing chemical modifications (selenophene, benzothiophene, benzofuran, fluorobenzofuran) at C5 position 2′-deoxyuridine serve suitable substrates for TdT, albeit differences incorporation efficiency. A battery experiments provided valuable insights into scope this functionalization method. It revealed how fine balance between steric hindrance stacking interaction heterocycle moiety primer 3′-end nucleobase active site modulates recognition processing based on their size. Remarkably, excellent responsiveness benzofuran-modified dUTP enabled design assays estimate activity, detect nucleotide non-nucleotide findings obtained using our should significantly advance TdT-based functionalization, diagnostic, therapeutic strategies.
Язык: Английский
Процитировано
0
International Journal of Biological Macromolecules, Год журнала: 2025, Номер unknown, С. 142712 - 142712
Опубликована: Март 1, 2025
Язык: Английский
Процитировано
0
Elsevier eBooks, Год журнала: 2025, Номер unknown
Опубликована: Янв. 1, 2025
Язык: Английский
Процитировано
0
Biotechnology Advances, Год журнала: 2025, Номер unknown, С. 108604 - 108604
Опубликована: Май 1, 2025
Язык: Английский
Процитировано
0
Chemical Science, Год журнала: 2024, Номер unknown
Опубликована: Дек. 18, 2024
Low steric hindrance, low error rate: conducting synthesis on a smooth surface made direct of 1000-nucleotide oligos reality.
Язык: Английский
Процитировано
2