Introduction
M6A methylation modification is a form of RNA modification widely existing in eukaryotic mRNA and even viral RNA. In 2011, the first true RNA demethylase FTO was reported, which made the methylation research of mrna/lncrna enter people's vision again. Merip was enriched and sequenced by m6A specific antibody for studying adenosine methylation modification of RNA.
Function introduction
Experimental strategy
Information analysis
Technical parameter
| Sample requirements | Project cycle |
| Total amount: different samples have different requirements, human samples ≥ 2 μ g; Sample type: total amount of complete total RNA samples after deproteinization and DNase treatment: no less than 70ug is recommended for routine samples, and the minimum sample concentration for trace methods can reach 8ug: it is recommended that ≥ 70 ng/ μ l sample integrity: Rin ≥ 7, ratio 28s/18s ≥ 0.7; Some source samples (such as body fluid samples, insect samples, aquatic organism samples, etc.) do not have RIN and ratio28s/18s requirements | The operation cycle of the standard process below 15 samples is about 50 working days. When the number of samples is large, the project cycle needs to be determined with the technical support personnel. |
Case analysis
Case：
Combined analysis of merip SEQ and RNA SEQ to study the molecular mechanism of heat shock response
Dynamic m6A mRNA methylation directs translational control of heat shock response
Background:
Heat shock response is a defensive adaptive response characterized by changes in gene expression, which is the body's response to heat stress. It is a non-specific emergency response in the face of stimuli to ensure that cells adapt to extreme environments temporarily. Heat shock protein (HSP) plays an important role in it. M6A is the most abundant mRNA epigenetic modification, regulating the variable splicing of transcripts, RNA protein interactions, and RNA stability. In mammalian cells, m6A is not symmetrically distributed, and its methylation level in the 5 'UTR of mRNA is lower than that in other functional regions. However, little is known about the mechanism of 5 'UTR methylation regulating gene expression, and 5' UTR plays a crucial role in the initiation of translation, but little is known about the regulatory role of m6A in translation.
Methods:
Result:
The relationship between m6A and heat shock response was studied, and the molecular mechanism of heat treatment affecting the methylation level of 5 'UTR of heat induced mRNA was mapped out: the nuclear localized m6A recognition protein ythdf2 inhibited the interaction of m6A demethylated protein FTO with the 5' UTR region of RNA, thus preserving the 5 'UTR methylation site of heat induced RNA. Studying the relationship between the dynamic changes of heat induced RNA 5 'UTR methylation and heat shock response not only expands the breadth of m6A regulation, but also reveals an unknown post transcriptional regulation mechanism of heat shock response, providing a new perspective for us to study the apparent modification.
