Discovery Of mRNA Research Articles

MRNA vaccination, a model of transition from basic biology to medicine

Sixty years elapsed between the discovery of messenger RNA (mRNA) and the use of this molecule in an unprecedented global vaccination campaign that brought the Covid-19 pandemic under control. Sixty years of doubts for some and certainties for others about the possibility of using mRNA-an example of synthetic biology-in therapeutic medicine and vaccinology. Years of "translational" research and development have culminated in the success of anti-Covid-19 mRNA vaccines and the promise of more to come against emerging pathogens. A new paradigm in vaccinology, enabling pandemics to be tackled as they emerge. A lesson to be learned: medical progress is less a question of time than of the critical nature of the biological discovery that underpins it. Before leaving us, François Gros, who played a key role in the discovery of mRNA, was able to appreciate the relevance of this obvious fact.

RNA-based vaccines – types, strategies of delivery and overview of RNA vaccines


 The discovery of mRNA by Sydney Brenner dates back to 1961, but the in vivo expression of mRNA was successful only in 1990, which initiated the development of vaccines based on this molecule. During Sars-CoV-2 pandemy the interest in the use of nucleic acids in the production of drugs and vaccines has increased significantly. The success of mRNA vaccines against Sars-CoV-2 has particularly empowered the pharmaceutical industry to create newer and newer generation products based on RNA modification that could help not only in Covid-2019, but also in the prevention and treatment of other infectious diseases. RNA has a very high potential - it can be used in highly personalized therapies, furthermore the production of mRNA is cheaper, faster than the current therapeutics and the process of mRNA making is more flexible due to the great ease of producing mRNA in the process of transcription. Modifying of the structure of ribonucleic acid and the methods of its delivery leads to the creation of newer and newer vaccines. In this review, we present the potential of RNA molecule in producing vaccines, types of RNA vaccines, strategies of RNA delivery and review of existing RNA-based vaccines.

T4 bacteriophage as a phage display platform.

Analysis of molecular events in T4-infected Escherichia coli has revealed some of the most important principles of biology, including relationships between structures of genes and their products, virus-induced acquisition of metabolic function, and morphogenesis of complex structures through sequential gene product interaction rather than sequential gene activation. T4 bacteriophages and related strains were applied in the first formulations of many fundamental biological concepts. These include the unambiguous recognition of nucleic acids as the genetic material, the definition of the gene by fine-structure mutation, recombinational and functional analyses, the demonstration that the genetic code is triplet, the discovery of mRNA, the importance of recombination and DNA replications, light-dependent and light-independent DNA repair mechanisms, restriction and modification of DNA, self-splicing of intron/exon arrangement in prokaryotes, translation bypassing and others. Bacteriophage T4 possesses unique features that make it a good tool for a multicomponent vaccine platform. Hoc/Soc-fused antigens can be assembled on the T4 capsid in vitro and in vivo. T4-based phage display combined with affinity chromatography can be applied as a new method for bacteriophage purification. The T4 phage display system can also be used as an attractive approach for cancer therapy. The data show the efficient display of both single and multiple HIV antigens on the phage T4 capsid and offer insights for designing novel particulate HIV or other vaccines that have not been demonstrated by other vector systems.

Switching from Prokaryotic Molecular Biology to Eukaryotic Molecular Biology

In the history of biology, the early 1950s to the mid-1960s represent a period of landmark discoveries that led to the establishment of a new science named Molecular Biology. The discovery of the double helical structure of DNA by Watson and Crick in 1953 facilitated the formulation of specific questions related to the hottest questions of the time, gene replication and expression. Many ambitious and talented scientists with varied backgrounds (geneticists, biochemists, microbiologists, physicists, and others) initiated work toward similar goals. Thus, starting with the confirmation in Escherichia coli of the hypothesis that DNA replication was semiconservative and, shortly thereafter, the discovery of mRNA, the formulation of the operon model for regulation of gene expression, and finally the elucidation of the genetic code, a flood of important discoveries arrived during this period. The favorite experimental system during this time was E. coli and its bacteriophages, i.e. prokaryotic systems. But, as exemplified by the demonstration of the universality (with minor exceptions) of the genetic code as well as the earlier demonstration of the similarity in major metabolic pathways such as glycolysis and the tricarboxylic acid cycle from bacteria to humans, biologists assumed that the fundamental biological principles discovered in E. coli must be largely correct for other organisms. In fact, there was a famous saying (generally attributed to Jacques Monod), “What is true for E. coli is also true for elephants.” Thus, there was a kind of feeling among ambitious people who participated in this early development of molecular biology that most of the important questions in biology, specifically questions asked using E. coli as a model organism, had been answered. Many people thought that cellular differentiation and morphogenesis could be the next major question in molecular biology and started to switch from E. coli to other experimental organisms, mostly eukaryotes, to study the questions related to differentiation or other phenomena uniquely observed in eukaryotes. The most extreme expression of this view came from Gunther Stent. Soon after the formal event marking the final decipherment of the genetic code, the Cold Harbor Symposium in 1966, he published an article in Science entitled “That Was the Molecular Biology That Was” (1), followed in 1969 by the publication of an expanded version as a book entitled “The Coming of the Golden Age: A View of the End of Progress” (2). After reviewing not only the history of progress in science, biology in particular, but also progress in other human activities, such as music and painting, Stent concluded that there may be no major conceptual breakthroughs remaining to be made in science. The exception perhaps would be in studying functions of the human brain, and this notion, the end of progress, might be equally true for art and other human activities. Such opinions were a strong influence on some early-day molecular biologists who had made significant contributions to the development of “classical molecular biology” using E. coli. This was exemplified by the exodus of many people from E. coli to other experimental systems at a time when many important problems in E. coli still remained unsolved. Like Seymour Benzer, who switched from E. coli molecular biology to Drosophila neurobiology in the late 1960s, Stent himself left E. coli molecular biology and switched to neurobiology using the leech as an experimental system. Among other early E. coli molecular biologists who switched their fields/systems, notably successful persons were, for example, George Streisinger, who initiated zebrafish research, and Sydney Brenner, who initiated research using the nematode worm Caenorhabditis elegans.

A Tribute to the Xenopus laevis Oocyte and Egg

When I was asked to reflect as part of the celebration of the 100-year anniversary of the Journal of Biological Chemistry I recalled a talk by Seymour Cohen at the Federation meetings in Atlantic City about 1960. He began by thanking bacteriophage for providing him with so many wonderful research problems. I decided in the same spirit to pay homage to the Xenopus laevis egg and oocyte, two states of a cell that has played and continues to play a central role in most of the disciplines of modern biology including biochemistry and molecular biology. Many of us owe a debt of gratitude to this cell. The egg is the most important and interesting cell in the repertoire of any organism. In X. laevis this single cell has a diameter of about 1.3 mm. The oocyte is permeable to small molecules, but after it undergoes meiosis and becomes an unfertilized egg it is impermeable to these same molecules. The oocyte is an active site of RNA and protein synthesis but not DNA replication, whereas the unfertilized egg is poised to replicate every 10 min after it is fertilized. During the cleavage period the embryo is transcriptionally silent. An oocyte can be cultured for days, but once ovulated the egg degenerates rapidly if it is not fertilized. The oocyte is the largest single cell in the body yet it has many of the same structures as somatic cells. These compartments are so exaggerated in size that they are accessible to cell biologists for manipulation and visualization. The huge oocyte nucleus is called a germinal vesicle (GV). The premeiotic tetraploid chromosomes are expanded into a “lampbrush” configuration so that they can be studied with a light microscope. The X. laevis egg and its possibilities for experimental manipulation were introduced to modern biology in the late 1950s by John Gurdon (Fig. 1), then a graduate student in the laboratory of Michail Fischberg in the Department of Zoology at Oxford University. Gurdon’s thesis was to reproduce with the South African “clawed toad” X. laevis the famous nuclear transplantation experiments that Briggs and King had perfected with the American leopard frog, Rana pipiens (1). Embryonic nuclei were injected into eggs whose own nuclei had been removed or destroyed. These early cloning experiments addressed the question of whether a differentiated and specialized somatic cell nucleus was still capable of expressing its full genetic repertoire. Briggs and King found that nuclei from R. pipiens embryos older than gastrula never promoted normal development, suggesting that the genetic material might change in cells as they specialize during embryonic development. This result was challenged by John Gurdon’s demonstration that nuclei from X. laevis embryos are more permissive. Even a small fraction of nuclei derived from differentiated intestinal epithelial cells supported normal development (2). This crucial scientific question of whether irreversible changes occur in the genome of highly specialized cells was addressed more precisely by experimental manipulation than by biochemistry or genetics. The concepts behind these experiments have influenced the stem cell field. The topic of whether the genome is altered in specialized somatic cells was revisited just this year by the same strategy of nuclear transplantation. Postmitotic nuclei from olfactory neurons support the development of normal fertile mice when transplanted into enucleated mouse eggs (3). THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 279, No. 44, Issue of October 29, pp. 45291–45299, 2004 © 2004 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A.

The discovery of mRNA.

The discovery of mRNA.

Regulation of ribosome biosynthesis in Escherichia coli and Saccharomyces cerevisiae: diversity and common principles.

In celebrating the centennial of the American Society for Microbiology, many people will surely recall the central importance that research using microbial systems played in the birth and the subsequent development of molecular biology in the latter half of the 100-year history. Starting from the demonstration of DNA as the genetic material, a series of key experiments, such as the proof of semiconservative replication of DNA, the discovery of mRNA as the information carrier between DNA and protein, and the eventual elucidation of the genetic code, were done mostly with microbial systems, the enteric bacterium Escherichia coli and its bacteriophages in particular. These basic principles in molecular genetics discovered with bacterial systems soon proved to be true for almost all organisms. Consequently, early research activities in molecular biology were concentrated on E. coli and related bacterial and phage systems, generating the initial attitude of many molecular biologists reflected in the well-publicized phrase, “What is true for E. coli is true for elephants.” (The acceptance of such an attitude at that time was not very surprising. Prior to the successful development of molecular biology, research in the field of intermediary metabolism from the 1920s through 1940s had demonstrated abundant evidence for the unity of biochemistry from microorganisms to humans, e.g., the mechanism of energy [ATP] production and its use for anabolic reactions [see also reference 42;]. Starting my first research as a student of fermentation biochemistry in 1950, I was certainly influenced by the prevalent belief, the unity of biochemistry, at that time.) Of course, in view of the bewildering diversity known in biology, especially some fundamental differences between prokaryotes and eukaryotes or single-cell versus multicellular organisms, such a view was expected to be too simple and naive. Thus, it was soon realized that the actual mechanisms and principles underlying certain biological functions, including diverse modes found in regulation of gene expression, are the consequences of evolutionary tinkering and may not necessarily be universal among diverse organisms (for a detailed discussion on evolution and tinkering, see reference 27). Nevertheless, attempts to extend factual observations or concepts obtained in one system (e.g., prokaryotes) to another (e.g., eukaryotes) have been made repeatedly and often turned out to be stimulating if not successful. As a person who was engaged in studies of synthesis of ribosomes and ribosomal components first in E. coli and later in Saccharomyces cerevisiae, I will recount some of the research activities on this subject which I have touched upon in this context.