Translation of the human genome into clinical allergy

ABSTRACTBy complete reading of the genome sequence, in the near future we will be able to determine the role of genomic DNA sequence variation among individuals, such a single nucleotide polymorphism (SNP), in the pathogenesis of diseases and responses to drugs. Comprehension of the genome will also accelerate understanding of the transcriptome, the whole transcripts present in a cell. Messages induced by a new therapy, such as an unexpected adverse effects, will not be missed by using such a comprehensive assay. Allergic diseases will be classified into subtypes depending on the impaired or affected molecule. Herein, I introduce our research strategy for genome-wide analysis of SNP related to asthma, granted by the Millennium Genome Project of the Japanese Government, and review the recent results of transcriptome analysis using microarray technology

Synthetic circular RNA switches and circuits that control protein expression in mammalian cells

mRNA医薬の課題克服に向けた「環状RNAスイッチ」の開発 --RNAによる持続的な遺伝子発現と細胞種特異的な制御に成功--. 京都大学プレスリリース. 2023-01-16.Cyclic RNA Switches that Regulate Gene Expression in a Cell Type-Specific Manner. 京都大学プレスリリース. 2023-02-16.Synthetic messenger RNA (mRNA) has been focused on as an emerging application for mRNA-based therapies and vaccinations. Recently, synthetic circular RNAs (circRNAs) have shown promise as a new class of synthetic mRNA that enables superior stability and persistent gene expression in cells. However, translational control of circRNA remained challenging. Here, we develop ‘circRNA switches’ capable of controlling protein expression from circRNA by sensing intracellular RNA or proteins. We designed microRNA (miRNA) and protein-responsive circRNA switches by inserting miRNA-binding or protein-binding sequences into untranslated regions (UTRs), or Coxsackievirus B3 Internal Ribosome Entry Site (CVB3 IRES), respectively. Engineered circRNAs efficiently expressed reporter proteins without inducing severe cell cytotoxicity and immunogenicity, and responded to target miRNAs or proteins, controlling translation levels from circRNA in a cell type-specific manner. Moreover, we constructed circRNA-based gene circuits that selectively activated translation by detecting endogenous miRNA, by connecting miRNA and protein-responsive circRNAs. The designed circRNA circuits performed better than the linear mRNA-based circuits in terms of persistent expression levels. Synthetic circRNA devices provide new insights into RNA engineering and have a potential for RNA synthetic biology and therapies

Target-dependent RNA polymerase as universal platform for gene expression control in response to intracellular molecules

抗体を用いた標的依存性RNAポリメラーゼの開発 多様な細胞内分子に応答する遺伝子発現制御のプラットフォーム. 京都大学プレスリリース. 2023-11-17.Using Single-antibodies as a New Ingenious Tool to Build Bio-circuitry. 京都大学プレスリリース. 2023-11-17.Controlling gene expression in response to specific molecules is an essential technique for regulating cellular functions. However, current platforms with transcription and translation regulators have a limited number of detectable molecules to induce gene expression. Here to address these issues, we present a Target-dependent RNA polymerase (TdRNAP) that can induce RNA transcription in response to the intracellular target specifically recognized by single antibody. By substituting the fused antibody, we demonstrate that TdRNAPs respond to a wide variety of molecules, including peptides, proteins, RNA, and small molecules, and produce desired transcripts in human cells. Furthermore, we show that multiple TdRNAPs can construct orthogonal and multilayer genetic circuits. Finally, we apply TdRNAP to achieve cell-specific genome editing that is autonomously triggered by detecting the target gene product. TdRNAP can expand the molecular variety for controlling gene expression and provide the genetic toolbox for bioengineering and future therapeutic applications

Versatile strategy using vaccinia virus-capping enzyme to synthesize functional 5′ cap-modified mRNAs

様々な5'キャップ構造をもつ機能的なmRNAの汎用的な合成方法 酵素を用いて簡便かつ効率的に. 京都大学プレスリリース. 2023-02-03.Development of a versatile method to synthesize functional mRNAs with diverse 5' cap structures. 京都大学プレスリリース. 2023-02-03.The potential of synthetic mRNA as a genetic carrier has increased its application in scientific fields. Because the 5′ cap regulates the stability and translational activity of mRNAs, there are concerted efforts to search for and synthesize chemically-modified 5′ caps that improve the functionality of mRNA. Here, we report an easy and efficient method to synthesize functional mRNAs by modifying multiple 5′ cap analogs using a vaccinia virus-capping enzyme. We show that this enzyme can introduce a variety of GTP analogs to the 5′ end of RNA to generate 5′ cap-modified mRNAs that exhibit different translation levels. Notably, some of these modified mRNAs improve translation efficiency and can be conjugated to chemical structures, further increasing their functionality. Our versatile method to generate 5′ cap-modified mRNAs will provide useful tools for RNA therapeutics and biological research

Factors associated with the severity of childhood rhinoconjunctivitis

AbstractBackgroundAllergic rhinitis is one of the most common chronic diseases in children. Although it has a large impact on the patient's quality of life, little is known about the factors associated with its severity. The aim of this study was to assess the factors associated with the severity of rhinoconjunctivitis among children in the general population.MethodsA survey was conducted using an online research panel in 2012. Parents were asked to answer an International Study of Asthma and Allergies in Childhood-based questionnaire to identify children with current rhinoconjunctivitis and evaluate factors associated with the severity of its symptoms. Severity was rated according to the degree of impairment caused by the symptoms in the patient's daily life.ResultsAmong 26,725 children aged 6–12 years old, rhinoconjunctivitis was defined in 5175 (19.4%), and of these, 688 children (13.3% of children with current rhinoconjunctivitis) presented severe symptoms. Living in areas with a high cedar and cypress pollen count and having concurrent eczema were associated with severe rhinoconjunctivitis [adjusted OR (95% CI): 1.21 (1.00–1.46) and 1.45 (1.20–1.75), respectively]. Further, a maternal history of asthma and allergic rhinitis was a significant risk factor for severe rhinoconjunctivitis [1.34 (1.04–1.74) and 1.30 (1.10–1.53), respectively]. However, living with fur-bearing animals (pets) before 1 year of age proved to be a protective factor against severe rhinoconjunctivitis [0.70 (0.52–0.94)].ConclusionsEnvironmental factors such as pets and pollen, together with comorbidities and a maternal history of allergic diseases, play an important role in determining the severity of rhinoconjunctivitis