Recently, RNA modifications located in the coding region of mRNAs have been regarded as critical regulators of gene expression. However, there are still many aspects related to the functions of these diverse mRNA modifications. We employ mass spectrometry (MS) to provide a comprehensive, accurate and affordable solution for mRNA modification analysis including mapping, quantitation and characterization.
Overview
RNAs undergo a variety of modifications, which is a reversible and dynamic process. Multiple chemical modifications, such as N6-methyladenosine (m6A), 5-methylcytosine (m5C), N1-methyladenosine (m1A) and pseudouridine (Ψ), have been detected within open reading frames (ORFs) of mRNAs. Among them, m6AThese modifications have been implicated with the fate of an mRNA, including its maturation, translation, and decay. For example, RNA m6A can direct mRNAs to distinct fates and accelerate mRNA metabolism and translation. MS offers a direct readout on any modifications that alter the mass of nucleoside and has long been a robust method for detecting, profiling, and characterizing known and unknown mRNA modification, profiling and characterizing modified mRNAs. Based on the LC-MS/MS platform, CD Genomics provides qualitative and quantitative information on all mRNA modifications under certain conditions or in diverse genetic backgrounds. We can help you detect the expression levels of as many as 36 types of modifications in a given sample (Table 1). Our mRNA modification analysis service by MS provides detailed insights into the function of mRNA modifications, mRNA biology and the development of targeted therapies.
Table 1. The list of mRNA modifications we have detected.
Nucleoside
Symbol
Nucleoside
Symbol
Nucleoside
Symbol
5-methylcytidine
m5C
5-hydroxymethylcytidine
hm5C
5,2'-O-dimethylcytidine
m5Cm
3-methylcytidine
m3C
1-methyladenosine
m1A
N6-methyladenosine
m6A
7-methylguanosine
m7G
1-methylguanosine
m1G
N2-methylguanosine
m2G
pseudouridine
Ψ
1-methylpseudouridine
m1Ψ
inosine
I
2'-O-methylcytidine
Cm
N6-isopentenyladenosine
i6A
3'-O-methyladenosine
3'-OMeA
N4-acetylcytidine
ac4C
2-thiocytidine
s2C
N2,N2,7-trimethylguanosine
m2,2,7G
3'-O-methylcytidine
3'-OMeC
2'-O-methyladenosine
Am
N4-acetyl-2'-O-methylcytidine
ac4Cm
4-thiouridine
s4U
2'-O-methyluridine
Um
N2,N2-dimethylguanosine
m22G
3'-O-methylinosine
3'-OMeI
2-thiouridine
s2U
5'-O-methylthymidine
5'-OMeT
5-methoxyuridine
mo5U
3'-O-methyluridine
3'-OMeU
2'-O-methylguanosine
Gm
3-methyluridine
m3U
3'-O-methylguanosine
3'-OMeG
5,2'-O-dimethyluridine
m5Um
5-methyluridine
m5U
2'-O-methylinosine
Im
5-methyl-2-thiouridine
m5s2U
Features
Experienced
Transcriptome-Wide
Bioinformatics Analysis
Quality Control
The MS experiment is performed by our experienced expert team.
Profile all mRNA modifications, either known or unknown.
Data QC; profiling of mRNA modifications; quantitative analysis; functional analysis
Quality control is executed following every procedure.
Project Workflow
1. Sample Preparation
RNA isolation from a cell line, tissue, or bacterial colony.
2. mRNA Isolation
Isolate mRNA molecules using sing oligo(dT) beads.
3. mRNA Digestion
Enzymatically digest RNAs into constituent ribonucleosides.
4. Mass Spectrometry
Quantitative measurement of modified ribonucleosides by liquid chromatography tandem mass spectrometry (LC-MS/MS).
Sample Requirements
We work with a wide range of sample types including protein solution, fresh tissue, cultured cells, blood, and microbial colonies. Please feel free to contact us for sample size.
Sample Storage: extracted protein should be stored at -80°C. Avoid repeated freezing and thawing.
Shipping Method:when shipping the sample, it is stored in a 1.5 mL Eppendorf tube, sealed with a sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Deliverable: data QC report, MS results, integrated experimental report (materials, methodologies, and bioinformatics analysis).
References:
Hoernes T P, Hüttenhofer A, Erlacher M D. mRNA modifications: Dynamic regulators of gene expression?. RNA biology, 2016, 13(9): 760-765.
Wetzel C, Limbach P A. Mass spectrometry of modified RNAs: recent developments. Analyst, 2016, 141(1): 16-23.
* For Research Use Only. Not for use in diagnostic procedures.
