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Related Concept Videos

Transfer RNA Synthesis02:36

Transfer RNA Synthesis

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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
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mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
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pre-mRNA Processing02:01

pre-mRNA Processing

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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
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MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy
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Therapeutic mRNA Engineering from Head to Tail.

Longfei Jia1, Shu-Bing Qian1

  • 1Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853, United States.

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Synthetic messenger RNA (mRNA) therapeutics offer advantages over DNA or protein-based options. Innovations enable programmable mRNA for diverse applications, though optimizing translation and stability remains a challenge.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Therapeutics

Background:

  • Synthetic messenger RNA (mRNA) can be translated into proteins, but instability and immune responses previously limited therapeutic use.
  • Technological advancements, including modified nucleosides, have transformed synthetic mRNAs into programmable therapeutic agents.
  • mRNA therapeutics offer advantages like flexible design, cost-effectiveness, and scalability compared to DNA or protein-based alternatives.

Purpose of the Study:

  • To review the general principles of mRNA translation in eukaryotic cells.
  • To elucidate the roles of coding and noncoding regions in translational regulation.
  • To highlight design strategies for engineering synthetic mRNAs with enhanced therapeutic potential.

Main Methods:

  • Discussion of fundamental principles of mRNA metabolism and translational control.
  • Analysis of sequence-dependent control of mRNA translation.
  • Focus on design strategies for synthetic mRNA components (5' cap, 5'UTR, CDS, stop codon, 3'UTR, poly(A) tail).

Main Results:

  • Synthetic mRNAs are versatile for encoding antigens, replacing defective proteins, or introducing novel functions like genome editing.
  • Recent successes in mRNA vaccines (e.g., against SARS-CoV-2) underscore the importance of understanding mRNA biology.
  • Massively parallel reporter assays (MPRA) are effective tools for identifying sequence elements that control mRNA translatability and stability.

Conclusions:

  • Rational design of synthetic mRNAs with optimal translation and stability is crucial for therapeutic development.
  • Understanding the interplay between sequence elements and translational output is key to advancing mRNA engineering.
  • This review aims to guide the rational design of synthetic mRNAs for broad therapeutic applications.