点击修饰的安纳胺siRNA可在神经元和免疫细胞中实现传递和基因沉默
Julian Willibald1, Johannes Harder, Konstantin Sparrer
1Center for Integrated Protein Science, Department of Chemistry, Ludwig-Maximilians-Universität , Munich, Germany.
Journal of the American Chemical Society
|July 21, 2012
概括
新的胺基改性RNA有效地将小干扰RNA (siRNA) 输送到具有挑战性的神经元和免疫细胞中. 这种新型RNA结合体显示出高的转染效率,并且无毒,扩大了基因沉默应用.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 药物运输 药物运输 药物运输
背景情况:
- 对于开发有针对性的基因沉默疗法,RNA修饰至关重要.
- 点击化学提供了一个多功能平台,用于将各种连接物与RNA结合起来.
- 开发小干扰RNA (siRNA) 的高效,无毒的输送系统仍然是一个挑战,特别是对于难以转移的细胞.
研究的目的:
- 使用点击化学合成新型RNA结合物.
- 评估安纳米德修饰RNA的转化效率和传递能力,用于siRNA.
- 在神经元和免疫细胞模型中评估安纳米德修饰RNA的潜力.
主要方法:
- 基因修饰RNA的合成.
- 点击化学结合与受体连接体亚化物 (叶酸,胆固醇,胺).
- 感染RBL-2H3 (神经元模型) 和BJAB (人体免疫) 细胞的siRNA载.
- 评估转染效率,基因沉默效应和细胞毒性.
主要成果:
- 对于3'-叶酸,3'-胆固醇和3'-胺基修饰RNA,获得了高产量和纯度.
- 胺基改性RNA表现出显著的转染性质,使siRNA能够传递到RBL-2H3细胞中.
- 在人类免疫BJAB细胞中证明了有效的基因沉默,与基准试剂相当.
- 发现安纳胺合物是无毒的.
结论:
- 点击化学提供了一个强大的方法来创建功能化的RNA.
- 胺基改性RNA代表了一个有希望的,非有毒的平台,用于siRNA传递.
- 这种方法扩大了基于siRNA的基因沉默在神经元和免疫细胞中的范围.
相关概念视频
02:30siRNA - Small Interfering RNAs
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
02:15Experimental RNAi
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...