相关实验视频
Updated: Jun 11, 2025

11:00
Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
Published on: June 12, 2018
13.7K
开发基于生物层干涉测量 (BLI) 的双链RNA检测方法,并应用于基于mRNA的疗法和疫苗
Dharia Sara Silas1, Bindiya Juneja1, Keerat Kaur2
1Protein Biochemistry, Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA.
Pharmaceutics
|September 28, 2024
概括
一种新的生物层干扰测量 (BLI) 试验能够快速而灵敏地检测体外转录 (IVT) mRNA中的双链RNA (dsRNA) 杂质,即使是尿素修饰. 这种方法通过在生产过程中监测dRNA去除,确保了更安全的mRNA治疗方法.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
背景情况:
- 在体外转录 (IVT) 的mRNA是治疗药物和疫苗的关键平台.
- 双链RNA (dsRNA) 是IVT mRNA中的关键杂质,可能导致不良免疫反应.
- 目前的dsRNA检测方法缺乏灵敏度,速度,或与修改后的尿素作斗争.
研究的目的:
- 调查已修改的dsRNA.现有方法的降低灵敏度.
- 开发一种用于IVTmRNA中dSRNA检测的快速和灵敏的测试方法.
- 为了在mRNA生产过程中实时监测dRNA杂质.
主要方法:
- 对抗dsRNA抗体的特征性结合亲和力,以了解敏感性限制.
- 开发了一种生物层干扰测量 (BLI) 试验,使用群病毒 (FHV) B2蛋白.
- 验证了BLI测定对各种dSRNA类型和修改的验证.
主要成果:
- BLI测定检测到具有尿素修饰 (ψ,m1ψ,5moU) 的dsRNA,具有高灵敏度.
- 量化短,长,和发针结构的dSRNA杂质.
- 在净化过程中成功监测了dsRNA去除.
结论:
- BLI测定提供了一种敏感和快速的dSRNA检测方法.
- 这种测试可以通过监测杂质来确保IVTmRNA的质量和安全.
- 能够实时监测杂质,以防止dRNA诱导的免疫性.
相关概念视频
Experimental RNAi
6.1K
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...
6.1K
RNA Interference
26.0K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.0K
Ribosome Profiling
3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K

