Capping Efficiency and Poly(A) Tail Characterization

Seattle Genova provides mRNA Analysis and Characterization through 5’ capping and 3’ polyadenylation tails.. The main goal of our services is to provide variable aspects of the transcript, which are critical contributes of mRNA stability and half-life. 

The five-prime cap (5′ cap) is a specially altered nucleotide on the 5′ end of some primary transcripts such as precursor messenger RNA. The 5′ cap has four main functions:

Regulation of nuclear export;

Prevention of degradation by exonucleases;

Promotion of translation (see ribosome and translation);

Promotion of 5′ proximal intron excision.

The primary purpose of the poly-A tail is to protect the newly synthesized mRNA from degradation. Moreover, the capping also assists in ribosome binding, which helps to initiate the translation and aids in exporting the mature mRNA to the cytoplasm and is involved in binding protein to begin the translation process.

To test the cap Efficiency and Poly(A) Tail Characterisation, Seattle Genova offers various techniques through which the efficiency of capping and poly-A tail characterization will be applied.

Capping Efficiency Analysis

For the multifunctional efficiency of mRNA, the 5' cap structure is capped and stabilized against exonucleolytic degradation. The multifunctional cap involves translation, splicing, stabilization, and nucleocytoplasmic transport. Due to the importance of the capping in mRNA, the orientation of capping is vital for understanding the efficiency analysis of mRNA translation.

Poly(A) Tail Length Analysis

Seattle Genova offers various lengths of poly(A) tail extended to an mRNA. This addition would determine the time of mRNA initial synthesis or remodeling post-synthesis through the concerted action of deadenylases and poly(A) polymerases. Hence, the poly(A) tail could be a dynamic entity whose length can significantly impact the biological fate of an mRNA transcript.

Biotin-tagged probes

Seattle Genova offers a label-free analysis of mRNA capping efficiency through biotin-tagged probes used with RNase H to cleave the 5' end of the mRNA. The cleaved end sequence was isolated through streptavidin-coated magnetic beads, and the capping efficiency could be determined between the range of 85-98%, depending on the modification and length of mRNA.

Ribozyme Assays

Seattle Genova designed a set of ribozyme assays that explicitly cleaves the IVT mRNA at a unique position and release 5' capped up to 30nt long. The capping efficiencies of IVT mRNA are analyzed through liquid chromatography and mass spectroscopy (LC-MS).

Deliverables

Seattle Genova provides in-depth mRNA Analyses and Characterization, which helps to produce precise and consistent results. Our services will provide reliable, accurate, and user-friendly results. Consequently, we believe in delivering results that manage customers' research with secure access.