Effects of Various Poly(A) Tails on Luciferase Expression

IF 0.6 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry Pub Date : 2024-11-05 DOI:10.1134/S1990750824600055
I. Kukushkin, O. Vasileva, D. Kunyk, S. Kolmykov, T. Sokolova, A. Muslimov, I. Terenin, O. Zaborova, A. Kirshina, F. Kolpakov, R. Ivanov, V. Reshetnikov
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Abstract

Synthesis of RNA by in vitro transcription has many applications in the production of vaccines, anticancer and gene therapy medicines. Despite several advantages of mRNA molecules over DNA in terms of transient transgene expression, rapid degradation of RNA is the limiting factor for the efficacy of mRNA-based therapeutics. Translational efficiency and stability of mRNA depends on the sequence of 5′ and 3′ untranslated regions, cap structure, coding sequence (CDS), and the poly(A) tail. So far, the role of the poly(A) tail sequence in mRNA stability and translational efficiency remains poorly investigated. There are few studies evaluating the influence of poly(A) tails containing certain non-A nucleotides or segments of non-A nucleotides on the efficacy of mRNA therapeutics. In our work, we conducted a comprehensive assessment of the effects of mRNA poly(A) tail on luciferase expression in cell lines HEK293 and DC2.4 and in BALB/c mice. Our results showed that mRNAs with a poly(A) tail consisting of 100 adenines separated by a segment of 10 nucleotides (50A-GCAUAUGACU-50A) have the best translational efficiency both in vitro (in cell lines) and in vivo (in BALB/c mice). Nonetheless, the results were inconsistent and depended both on cell lines and on the coding sequence (NanoLuc or firefly luciferase). Thus, effects of a mRNA poly(A) tail on luciferase expression may depend on the sequence of the gene in question and on specific features of the expression of RNA-binding proteins in cell lines.

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各种聚(A)尾对荧光素酶表达的影响
通过体外转录合成 RNA 在生产疫苗、抗癌药物和基因治疗药物方面有许多应用。尽管与 DNA 相比,mRNA 分子在瞬时转基因表达方面具有一些优势,但 RNA 的快速降解是限制基于 mRNA 的疗法疗效的因素。mRNA 的翻译效率和稳定性取决于 5′和 3′非翻译区的序列、帽结构、编码序列(CDS)和多聚(A)尾。迄今为止,关于多聚(A)尾序列在 mRNA 稳定性和翻译效率中的作用的研究仍然很少。很少有研究评估含有某些非 A 核苷酸或非 A 核苷酸片段的聚(A)尾对 mRNA 治疗效果的影响。在我们的工作中,我们全面评估了 mRNA 多(A)尾对 HEK293 和 DC2.4 细胞系以及 BALB/c 小鼠荧光素酶表达的影响。我们的结果表明,mRNA 的聚(A)尾由 100 个腺嘌呤组成,腺嘌呤之间相隔 10 个核苷酸(50A-GCAUAUGACU-50A),在体外(细胞系)和体内(BALB/c 小鼠)的翻译效率最高。然而,结果并不一致,而且取决于细胞系和编码序列(NanoLuc 或萤火虫荧光素酶)。因此,mRNA 聚(A)尾对荧光素酶表达的影响可能取决于相关基因的序列以及细胞系中 RNA 结合蛋白表达的具体特征。
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来源期刊
CiteScore
1.10
自引率
0.00%
发文量
31
期刊介绍: Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry   covers all major aspects of biomedical chemistry and related areas, including proteomics and molecular biology of (patho)physiological processes, biochemistry, neurochemistry, immunochemistry and clinical chemistry, bioinformatics, gene therapy, drug design and delivery, biochemical pharmacology, introduction and advertisement of new (biochemical) methods into experimental and clinical medicine. The journal also publishes review articles. All issues of the journal usually contain solicited reviews.
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