使用RNA拉下方法在小鼠皮质中研究体内circRNA相互作用的协议
Valentina Silenzi1, Nicolò Salvi1, Irene Bozzoni2
1Department of Biology and Biotechnologies "Charles Darwin," Sapienza University of Rome, 00185 Rome, Italy.
STAR protocols
|September 19, 2025
概括
这项研究详细介绍了一项新协议,用于识别小鼠大脑组织中的循环RNA (circRNA) 相互作用体. 该方法使用紫外线交联和RNA拉下来寻找与特定circRNAs相关的蛋白质.
科学领域:
- 分子生物学分子生物学
- 神经科学是一个神经科学.
- 基因组学就是基因组学.
背景情况:
- 循环RNAs (circRNAs) 是一种非编码RNAs类,在基因调节中扮演着新兴的角色.
- 识别circRNA结合蛋白对于理解它们的功能至关重要.
- 现有的方法可能在保护本地相互作用方面存在局限性.
研究的目的:
- 为在小鼠皮层中识别circRNAs的蛋白相互作用体提供一个强大的协议.
- 优化保存本地circRNA-蛋白质复合体的技术.
- 为神经系统研究提供一种可适应的方法.
主要方法:
- 小鼠皮质组织的组织解离和紫外线交联,以稳定RNA-蛋白相互作用.
- 用特定的圆形RNA (circDlc1(2) 作为诱进行RNA拉下测试.
- 优化步骤,以有效地分离与circRNA相关的分子.
主要成果:
- 该协议成功识别了与circDlc1(2) 相关的潜在蛋白相互作用体.
- 紫外线交联有效地维持circRNAs和蛋白质之间的本地相互作用.
- 该方法在小鼠大脑组织中被证明是有效的.
结论:
- 该协议为发现中枢神经系统中circRNA-蛋白相互作用提供了一种可靠的方法.
- 该技术可以适应用于研究其他circRNA和大脑区域.
- 进一步的研究可以利用这个协议来阐明神经过程中的circRNA功能.
相关概念视频
RNA Structure
Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
RNA Interference
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...
RNA Interference
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...
siRNA - 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...
Experimental 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...
Small interfering RNAs (siRNA)
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...


