Gene is the third wave of revolution in the industry, and it has entered the era of rapid growth. According to Frost &Sullivan**, the global CGT market space is expected to grow from 20$800 million to $305 by 2025400 million US dollars, with a compound growth rate of 711%。Among them, China will be the fastest growing market, from 200 million US dollars in 2020 to 1.7 billion US dollars in 2025, with a compound growth rate of 511%。In the face of such a gradually huge market, the production of vectors A** and lentivirus, which are the most commonly used in the field of genetics, require plasmids as important pharmaceutical raw materials, so a large number of plasmids that meet the quality requirements are needed every year to meet the market demand in the field of genetics.
Adeno-associated virus belongs to the family Parvoviridae and is a defective virus with a simple structure and non-pathogenic. Adeno-associated viruses are replication-deficient and often require a helper virus, such as adenovirus or herpesvirus, to replicate themselves during their life cycle. The production of adenoviral vectors is often cultured and expanded in specific cell lines, and high infectivity and virulence can lead to cell death, so maintaining proper cell culture conditions is critical for efficient virus production. For large-scale production of adenoviral vectors, such as genetic clinical trials or the production of vaccines, scalable production processes need to be established to ensure adequate yield and consistent product quality. Lentiviruses are essentially a class of retroviruses, and compared to other retrovirus types, lentiviruses are able to penetrate the nuclear envelope, infect a wider range of cell stages, and achieve efficient infection of both ** cells and non** cells. Lentiviral vectors (LVs) are currently the most mature type of viral vectors in the industry, especially in immune cells**, such as CAR-T**. However, due to the instability of physicochemical properties and the low rate of lentiviral vectors, the development and optimization of downstream purification processes is the bottleneck and key to the large-scale production of lentivirus. In addition, the production cost of lentiviral vectors mainly comes from upstream disposable consumables and plasmid transfection systems, so the development of large-scale production and the development of stable cell culture systems that do not rely on transfection systems are the key to reducing costs. The global demand for DNA is increasing, and it is likely to rise from 3kg per year to 10kg or even 100kg per year. Experts began to worry that the way DNA was produced would become a bottleneck, and the timely advent of end-blocked linear DNA solved the immediate need. While traditional antibodies** and plasmid DNAs** have long production cycles and high costs for products or plasmids, end-blocked linear DNA is the smallest, double-stranded, and covalently closed DNA vector amplified by a double-enzyme process in vitro, completely free of any bacterial propagation elements and antibiotics. It has been proven to be used in the production of viral vectors, cells**, and DNA vaccines as an alternative to plasmids. The end-blocked linear DNA that can be used as a DNA carrier is based on the in vitro DNA enzymatic synthesis system, which perfectly avoids many uncontrollable risks in the biological fermentation process, and can serve many emerging technologies and has a wide range of application scenarios. Gene synthesis and modificationTerminally blocked linear DNA can be used for DNA sequences required for synthetic genes, genetic routes, or synthetic biology engineering projects. This is important for the development of new biological functions and the modification of microorganisms to produce specific compounds. Vaccine and drug developmentVaccines and drugs require a large number of DNA sequences, and the synthesis of end-blocked linear DNA can be used to construct expression vectors expressing antigens or drug-related proteins to accelerate related research. Gene editing and CRISPR-Cas9 technologyIn gene editing, end-blocked linear DNA can be used to provide repair templates or targeted primers for gene editing, insertion, or deletion. In addition, it helps to improve the efficiency of CRISPR-Cas9 technology. Synthetic Biology ProjectSynthetic biology projects often require large-scale DNA synthesis. Terminally blocked linear DNA can be used to construct components and components for synthetic biology engineering projects. Gene regulation studiesResearchers can synthesize gene regulatory elements, such as promoters, operons, and regulatory elements, using terminal-blocked linear DNA to delve into gene regulatory networks. Disease diagnosis and testingIn the field of molecular diagnostics, end-blocking linear DNA can be used to design primers and probes for disease diagnosis and detection. DNA barcodeIn the field of biodiversity research and DNA barcoding, end-enclosed linear DNA can be used to generate DNA barcode sequences to identify different biological species. Protein expression and analysisProtein expression and analysis studies that require a large number of DNA sequences, such as the construction of expression vectors and prokaryotic or eukaryotic expression systems. Custom DNA sequencesEnd-blocking linear DNA technology enables the synthesis of customized DNA sequences for specific needs for a variety of biological research and engineering applications. Genetic engineeringEnd-blocking linear DNA can be used to construct engineered microorganisms or cells to produce products such as compounds, biomaterials, or biofuels. These application areas highlight the importance of end-blocking linear DNA technology in life sciences and bioengineering. It provides researchers and engineers with an efficient and customizable method for creating specific DNA sequences, driving many innovative projects and research. High-precision synthesisEnd-blocking linear DNA technology enables precise synthesis of specific DNA sequences without the need for complex cloning steps. This avoids the introduction of errors or mutations and ensures the high accuracy of the required DNA sequences. EfficiencyCompared with traditional DNA synthesis methods, end-blocking linear DNA technology is more efficient. It allows for large-scale DNA synthesis and helps accelerate the progress of research and engineering projects. CustomizabilityEnd-blocking linear DNA can be customized to meet the needs of a research or engineering project. Researchers can choose the desired DNA sequence, including specific genes, promoters, primers, and more. ScalabilityEnd-blocking linear DNA technology is scalable and suitable for large-scale DNA synthesis projects. This is useful for synthetic biology engineering projects and the production of compounds. Overall, the precision, efficiency, customization, and versatility of end-blocked linear DNA technology make it a powerful tool in the fields of synthetic biology and molecular biology, helping to accelerate research and application progress. It helps unlock the potential of many life sciences and bioengineering projects to drive innovation and discovery. Prosperity Technology is sharing with you today the article published in Human Vaccines & Immunotherapeutics: novel synthetic plasmid and doggybone TM DNA vaccines induce neutralizing antibodies and provide protection from lethal in uenza challenge in mice.This article focuses on nucleic acid vaccines (n**s) that increase the availability of influenza vaccines due to their simple design and fast production. N**S can also target multiple influenza antigens and control influenza variation. The article generates the delivery of linear DNA cassettes through an enzymatic process, including promoters, DNA antigens, polya tails, and antigen expression cassettes at the end, which have many advantages over plasmid DNA. The specific response of CD4 and CD8 T cells was also characterized, the dCBDNATM-induced hemagglutination inhibition (HI) titer was measured and compared to the response of an optimized plasmid DNA (PDNA) vaccine encoding the same H1N1 influenza A PR 8 34 Ha gene, yielding similar humoral and cellular immune responses. Both structures induce high titers of Hi antibodies, protecting animals from deadly viruses. fig.Construction and representative expression of dbDNATM PR8 and pDNA PR8 constructs. (a) The process of enzymatic production of dbdnatm. Rolling circle amplification of the double-stranded DNA template yields linkers that are cleaved and attached by the telomeric preenzyme teln, resulting in a covalently closed double-stranded cassette. (b) Schematic diagram of the structure of a linear double-stranded dbdnatm pr8 with terminal single-stranded DNA hairpins. The final product removes the plasmid backbone sequence with restriction enzymes and exonucleases. (c) Schematic diagram of the structure of PDNA PR8, PR8 ha . ecro。The PR8 sequence was cloned into the PVAx1 mammalian expression vector. CMV promoter, HA gene, BGH poly-A signal, kanamycin resistance gene, and PUC origin are shown. (D) In vitro expression of representative DBDNATM PR8 and PDNA PR8 constructs. Confirm expression with transfected RD cells and HA-labeled antibodies. Empty vector (PVAX) as a negative control. The results were analyzed with confocal imaging. Fluorescein isothiocyanate (FITC) staining (green) indicates expression. HongXun Technology has a complete plasmid preparation production line to ensure that it provides high-quality sterile plasmids without RNA contamination and genomic contamination, and can control the endotoxin content at a low level (< 100eu mg, <30eu mg, <5eu mg, optional). In addition, we also provide sequence verification, restriction enzyme digestion and endotoxin analysis services to meet the diverse downstream applications of customers in various fields, such as transfection, antibody production, vaccine and gene research, etc. Terminally blocked linear DNA has advantages in applications that require high precision, efficiency, and customization, while plasmids may be even more advantageous in traditional molecular biology research and applications with high stability requirements. Our experts will select the appropriate DNA material according to the specific needs of the project. veronical scott, novel synthetic plasmid and doggybone tm dna vaccines induce neutralizing antibodies and provide protection from lethal influenza challenge in mice.walters, a., kinnear, e., shattock, r. et al. comparative analysis of enzymatically produced novel linear dna constructs with plasmids for use as dna vaccines. gene ther 21, 645–652 (2014).m.barreira, c.kerridge, s.jorda, et al., enzymatically amplified linear dbdnatm as a rapid and scalable solution to industrial lentiviral vector manufacturing. gene therapy, 2023.