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循环RNA过度表达的改进模型:使用Actin Intron揭示了高循环化效率

Feiya Li^1,2, Juanjuan Lyu^1,2, Yang Yang^1,2

¹Sunnybrook Research Institute Sunnybrook Health Sciences Centre Toronto ON M4N 3M5 Canada.

Advanced genetics (Hoboken, N.J.)

|September 28, 2023

概括

一种新的方法提高了循环RNA (circRNA) 表达效率,使用actin内序列. 优化的内子长度,特别是60-100个核酸,显著改善了用于功能研究的circRNA生产.

科学领域:

分子生物学分子生物学
在RNA生物学,RNA生物学.
生物技术是生物技术.

背景情况:

使用T4 td基因组1内子进行循环序列生成的传统方法表现出低循环RNA (circRNA) 表达效率.
低效率阻碍了对circRNAs的强有力的功能研究.

研究的目的:

开发一种新的,高效的方法来表达循环序列,特别是circRNAs.
通过测试不同长度的actin intron序列来优化circRNA表达.

主要方法:

使用actin内序列 (15-nt至180-nt) 和T4内构建circRNA表达等离子体.
在人 (293T) 和小鼠 (B16) 细胞系中引入等离子体.
通过连接检测,测序和RNA拉下测试,然后通过质谱 (MS) 评估循环化效率.

主要成果:

中长的actin内子 (60-nt至100-nt) 显示循环化效率显著提高.
在大多数测试条件下,100nT的actin内子表现出最高的效率.
质谱测量确定了参与高循环化机制的潜在拼接因素.

结论:

一个新的circRNA表达系统利用优化的actin内质长度提供了高循环化效率.
这种改进的系统为未来的circRNA功能研究提供了更可靠的平台.
这些发现有助于推进研究circRNA及其生物学作用.

关键词:

在T4内中.在Actin Intron中使用了Actin.环RNA 环RNA 是一个环RNA.圆形的RNARNA是一个圆形的RNA.过度表达是一种过度表达.这是一个拼接因子.

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