SCREENING METHOD AND SYSTEM FOR POSTTRANSCRIPTIONAL REGULATORY AGENT

• Main Authors: YAMASHITA AKIO, ONO SHIGEO
• Format: Patent
• Language: English; Japanese
• Published: 25.01.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; VINEGAR; WINE

To provide a screening method for post-transcriptional regulatory agents that enables accurate and highly efficient screening for post-transcriptional regulatory agents, which regulate nonsense mutation-dependent mRNA degradation (NMD) and post-transcriptional degradation of miRNA, and does not require long treatment with candidate compounds; and to provide a system therefor.SOLUTION: A screening method for post-transcriptional regulatory agents comprises: treating with a candidate compound for a transcription inducer, a read-through agent, and a post-transcriptional regulatory agent a cell containing a nucleic acid construct containing an inducible promoter and a first reporter gene in which a premature stop codon has been introduced into an exon, which is functionally linked to the promoter; and selecting as a post-transcriptional regulatory agent a candidate compound with detected reporter activity.SELECTED DRAWING: Figure 1A 【課題】ナンセンス変異異存mRNA分解(NMD)やmiRNAの転写後分解を制御する、転写後制御剤を的確に、かつ、高効率にスクリーニングすることができ、候補化合物による長時間の処理を必要としない、転写後制御剤のスクリーニング方法及びそのためのシステムを提供すること。【解決手段】転写後制御剤のスクリーニング方法は、誘導性プロモーターと、該プロモーターに機能的に連結された、エキソン内に未成熟終止コドンが導入された第１のレポーター遺伝子とを含む核酸構築物を含む細胞を、転写誘導剤、リードスルー剤及び転写後制御剤の候補化合物で処理し、レポーター活性が検出された候補化合物を転写後制御剤として選択することを含む。【選択図】図１Ａ