tRNA with s²U modifications shows a more stable structure and resistance against extreme thermophiles. Lack of the s²U modifications will result in growth defects, sensitivity to high temperatures, even protein misfolding in bacteria. Abnormality of s²U modification in humans is implicated with acute infantile liver failure and other dysfunctions.

https://mrna.creative-biolabs.com/2-thiouridine.htm

5-Methylcytidine (m⁵C), also called 5-methyl-cytidine, is a modified nucleoside derived from 5-methylcytosine (5mC), in which a methyl group is linked to the C-5 in the 6-atom ring of cytosine base catalyzed by DNA methyltransferase. 5mC has been considered to be the fifth DNA base, which is common in DNA deoxynucleotides of many animals and plants, also is found in ribonucleic acids of animal, plant, and bacterial organisms. In animals, especially vertebrates and mammalian, 5mC is mainly present in cytosine-guanine sequence (CpG), while, 5mC can be found in multiple sites and served as a marker to protect DNA from being cut by the methylation-sensitive restriction endonuclease. In vitro, 5mC can be derivatives into 5-hydroxymethylcytosine and thymine under corresponding reaction conditions.

https://mrna.creative-biolabs.com/5-methylcytidine.htm

The classical cap structure is a 7-methylguanosine moiety connected to the mRNA via a triphosphate linkage at 5'-end (m7GpppG or m7G). It can be recognized by several specialized proteins involved in mRNA transport, translation, translational repression and degradation. 6-thioguanosine (s6G) is a photosensitive molecule that can photo-crosslink to both proteins and nucleic acids to study cap-protein and cap-nucleic acid interactions at the molecular level.https://mrna.creative-biolabs.com/6-thioguanosine-containing-cap.htm

Methods of mRNA stability tests such as transcription inhibition initially are proved most amenable for high-throughput technologies. Actinomycin D is a fast-acting DNA intercalator which is widely used as a transcription inhibitor to achieve mRNA inhibition in mRNA stability assays. The level of an mRNA transcript could be quantified to measure the half-life of specific mRNAs either when the mRNA is in a steady-state or after the inhibition of mRNA synthesis by actinomycin D.

https://mrna.creative-biolabs.com/actinomycin-d-based-method.htm

Actinomycin D-based method as an easy approach has several advantages, for example, it does not require the construction and introduction of exogenous genes into cells. Although this method does not permit quantitative estimates of mRNA stability, it allows the easy analysis of mutant genes and thus may be useful for measuring the mRNA stability mechanism. Moreover, actinomycin D-based methods can determine the abundance of mRNA sequences following transcription inhibition, which allows the assessment of mRNA decay and stability.

https://mrna.creative-biolabs.com/actinomycin-d-based-method.htm

Methods of mRNA stability tests such as transcription inhibition initially are proved most amenable for high-throughput technologies. Actinomycin D is a fast-acting DNA intercalator which is widely used as a transcription inhibitor to achieve mRNA inhibition in mRNA stability assays. The level of an mRNA transcript could be quantified to measure the half-life of specific mRNAs either when the mRNA is in a steady-state or after the inhibition of mRNA synthesis by actinomycin D.

https://mrna.creative-biolabs.com/actinomycin-d-based-method.htm

Messenger ribonucleic acid (mRNA), a type of RNA molecule that can be found in various cells. In general, the structure of mRNA is often composed of a 5' cap, two untranslated regions (5' UTR and 3' UTR), a coding domain, a 3' poly(A) tail. mRNA can be produced during the process of transcription in the nucleus. Normally, primary transcript mRNA (pre-mRNA) will be created at the early stage of transcription. This pre-mRNA will be further processed into mature mRNA by RNA splicing and removing introns. Mature mRNA can be further transported into the cytoplasm to participate in protein synthesis. Moreover, two main types of RNA molecules, transfer RNA (tRNA) and ribosomal RNA (rRNA), have also been identified that play a critical role in translating codons into amino acids in the protein-manufacturing process.https://mrna.creative-biolabs.com/applications-of-mrna-as-research-tools.htm

The mRNA 5ʹ cap regulates splicing, nuclear output, resistance to exonuclease degradation, and initiation of translation. In short, 5' cap has the following functions:

Regulation of nuclear export;

Prevention of degradation by exonucleases;

Promotion of translation;

Promotion of 5' proximal intron excision.

https://mrna.creative-biolabs.com/mrna-5-capping.htm

With the development of IVT methods, synthetic mRNA has become an emerging class of gene therapy.https://mrna.creative-biolabs.com/custom-ivt-synthesis-of-mrna.htm

Non-integrative mRNA reprogramming has proven to be the most efficient and clinically-relevant method for human induced pluripotent stem cells (iPSCs) reprogramming.https://mrna.creative-biolabs.com/custom-ips-cell-reprogramming-by-mrna.htm