ChIRP和CHART法的原理都是基于甲醛交联法,研究RNA与蛋白的相互作用。 UV光则以零距离和不可逆的共价键形式促进蛋白质与核酸发生交联,有轻度的尿苷交联倾向性,而不会促进蛋白质—蛋白质交联,这点比甲醛交联法更特异,但对于双链RNA的交联效果偏差,对于蛋白的交联效果取决于氨基酸。RAP、PAIR、MS
最近研究发现数百种新RBPs的RNA结合域尚不清楚,于是强调RNA蛋白复合物的复杂性和多样性,进而最新研究成果扩展了RNA-protein 相互作用的两大类方法:一类方法是以RNA为中心,研究与感兴趣的RNA结合的protein;另一类方法是以蛋白质为中心,研究与感兴趣的protein结合的RNAs。由于每种方法都有其优势和局限,选择最佳方法来...
The RNA ChIP-IT® Kit was designed to study RNA-protein interactions in a chromatin context, and has been optimized to recover RNA for RT-PCR analysis. The kit contains sufficient material for 25 assays, with optimized protocols using magnetic beads, to minimize labor required to obtain ...
AATF interacts with coding and non-coding RNA species. (A) Pie chart depicting the distribution of RNA biotypes bound by AATF other than rRNA. 69% of transcripts other than rRNA bound by AATF are protein-coding, (14%) snoRNA and scaRNA, and “non-coding RNA” biotypes (17%) encompass...
little is known about what determines RNA accumulation in condensates and to which extend single condensates differ in their RNA composition. To address this, we developed an approach to read the RNA content from single synthetic and protein-based condensates using high-throughput sequencing. We find...
To examine the RNA-binding activity of HARD protein, we expressed and purified the recombinant HARD protein fused with EGFP fromE. coli(Fig.1b). As a control, we expressed and purified EGFP in the same manner. The HARD protein adopts a stable monomeric structure as shown by gel filtration ...
While our workflow is similar to other protocols published in the literature and/or proposed by commercial reagent providers, we have introduced a key modification that addresses efficiently the most challenging step of the protein isolation process: the solubilization of the protein pellet after RNA ...
In addition to the three traditional types of RNAs (rRNAs, tRNAs, and mRNAs) that are directly involved in protein synthesis, many types of small non-coding RNAs (<200 nt) have been identified to participate in various biological processes: this includes snRNAs in pre-mRNA splicing, snoRNAs...
(ASS1) expression, also leads to increased cytosolic fumarate levels and fumarate-mediated protein succination. Pharmacological inhibition and genetic ablation of the tricarboxylic acid cycle enzyme fumarate hydratase (FH) further increases intracellular fumarate levels. Mitochondrial respiration is also ...
2.4. Computational tools used for prediction of RNA structures, ORFs, IRES, m6A sites, protein structures, and domain analysis The circRNA sequences used in the study were taken from the CIRCexplorer2 analysis of HeLa cell RNA-seq data as described above. To find the known circRNAs in our ...