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、MS2- BioTRAP和TRIP法的...
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 ...
The ubiquity of RNA-seq has led to many methods that use RNA-seq data to analyze variations in RNA splicing. However, available methods are not well suited for handling heterogeneous and large datasets. Such datasets scale to thousands of samples across dozens of experimental conditions, exhibit ...
We validate the LIM-domain-only protein Csrp1 as a tissue-dependent RNA binding protein. Taken together, HARD-AP is a powerful approach that can be used to identify RBPomes from any type of sample, allowing comprehensive and physiologically relevant networks of RNA-protein interactions. Similar ...
MiR-21 regulates purine metabolism leading to a DTP state Previous studies showed that miR-21-5p induced drug resistance to the 1st generation of EGFR-TKI gefitinib through decreasing expression of tumor suppressor phosphatase and tensin homolog (PTEN) or programmed cell death protein 4 (PDCD4) in...
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...
We next performed multiplex IHC to assess coexpression of TIGIT + on CD8 + T cells and PVR + on CK19 + cancer cells at the protein level. We detected both cell types expressing the ligand-receptor pair in close proximity to each other, supporting the potential for ...
(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 ...
To preserve intact a significant fraction of the cells during the metabolite extraction step, and to lyse them after the metabolite extraction, aids in obtaining a protein fraction for proteomics that includes such hydrophobic proteins. Fourthly, to perform the first mechanical lysis step at very ...