Methods of Evaluation in Orthopaedic Animal Research
Yuehuei H. An, Richard J. Friedman in Animal Models in Orthopaedic Research, 2020
The basic terminology given here is adapted from the reviews by Shore and Kaplan.251,252 A gene is a unit of heredity, consisting of a segment of chromosomal DNA that is required for production of a functional protein or RNA. The gene contains both coding and regulatory regions. A transgene is a foreign gene which has been spliced into an animals original genomic DNA. mRNA is a type of RNA that contains protein coding information. Nucleotide sequence refers to the order of nucleotides in a given segment of DNA or RNA. Translocation is the transfer of a portion of DNA from one chromosome to another. A probe is a DNA or RNA molecule that is labeled, or tagged, and can then be used to locate a complementary DNA or RNA strand through hybridization. Vectors are DNA molecules that are used as carrier molecules for cloned DNA sequences. They contain information which allows recombinant molecules to be replicated in host bacterial cells. A plasmid is a small circular double-stranded DNA molecule which is found in bacteria and replicates independently of the host chromosome. They are commonly used as vectors in molecular cloning. A recombinant DNA molecule is a DNA molecule containing segments of DNA from different origins, such as a piece of human DNA that has been joined to a plasmid DNA. A clone is a term used to describe identical segmental DNA molecules produced by recombinant DNA technique. Molecular cloning is a process by which a specific segment of DNA is isolated and then numerous identical copies, or clones, of that segment of DNA are generated.
Marvellous molecules
Brendan Curran in A Terrible Beauty is Born, 2020
Recombinant DNA technology is particularly good at producing proteins whose therapeutic value depends on their availability in the bloodstream. However, there is not a lot that this type of therapy can do for thousands of other conditions in which the defective proteins are to be found inside the cells; these include sickle cell anaemia, cystic fibrosis, Huntington’s chorea, many cancers and a multitude of other conditions. Recombinant DNA technology could, of course, be used to make lots of the missing protein in a fermenter but it would be impossible to get the product into the appropriate cells of the patient at the proper time and in the correct amounts. Faced with the fact that they cannot transplant functional proteins into cells, genetic engineers are attempting to rectify these conditions by a much subtler scheme. It is so powerful that, if successful, it would be able to cure not only those scourges listed a few lines ago but make even the exciting technologies described in this chapter very rapidly redundant.
Protein Subunit Vaccines and Recombinant DNA Technology
F. Y. Liew in Vaccination Strategies of Tropical Diseases, 2017
The essential feature of recombinant DNA technology or genetic engineering is the in vitro manipulation of foreign DNA such that it can be stably introduced into a suitable host cell or organism. This is followed by amplification of the gene or genes, followed by expression of the gene encoded by the foreign DNA to produce a so-called recombinant protein. Depending on the subsequent requirements of the system the protein is then extracted, purified, and tested for functional activity. DNA techniques can be broken down into a number of defined areas such as enzyme analysis, hybridization, identification of recombinant clones, vectors, hosts, transformation, directed mutagenesis, expression, extraction, and purification.
The potential of plant-made vaccines to fight picornavirus
Published in Expert Review of Vaccines, 2020
Omayra C. Bolaños-Martínez, Sergio Rosales-Mendoza
The plant-based vaccines must follow the guidelines defined by the FDA and the US Department of Agriculture that establish the evaluation of: 1) the presence of allergenic or toxic compounds, 2) the method of the plant production and propagation, 3) the characterization of the recombinant DNA, and 4) genetic stability for those cases based on stable transformation events [64]. On the other hand, environmental concerns should be taken into consideration to control the spread of the bioengineered pharmaceutical plants and meet regulations for global commercialization. Moreover, as for any technology, social acceptance is a critical factor and in the case of technologies involving GMOs; robust programs to make the society aware of the benefits and the minimal risks involved with this approach are crucial [65].
Allergic rhinitis management: what’s next?
Published in Expert Review of Clinical Immunology, 2018
Farnaz Tabatabaian, Thomas B Casale
Other novel therapies with modified extracts that are under investigation include recombinant vaccines and allergoids. Similar to peptide therapy, recombinant vaccines and allergoids have reduced allergenicity while preserving immunogencity. Recombinant vaccines are reproduced via recombinant DNA technology. There are two main strategies of development for recombinant allergens: either they are adjusted variants of natural allergens or are molecules that mimic the properties of natural allergens. The vaccines have been shown to be effective and safe [30]. However, more studies are needed to evaluate this form of therapy. Allergoids are chemically modified allergens via treatment with glutaraldehyde or formaldehyde. This chemical modification results in decreased IgE epitopes while preserving the T-cell epitopes. Allergoids are widely used in Europe. Allergoids allow faster up dosing and improved safety profile making them an attractive alternative for patients with severe allergic reactions to SCIT.
Immunotoxins and nanobody-based immunotoxins: review and update
Published in Journal of Drug Targeting, 2021
Mohammad Reza Khirehgesh, Jafar Sharifi, Fatemeh Safari, Bahman Akbari
ITs are new tools for cancer therapy that consists of two functional components: targeting and cytotoxic moieties. In ITs design, the binding domain of the protein toxin, responsible for binding to a specific receptor, replaces with a targeting moiety, usually mAbs [17]. Therefore, non-protein toxins such as Brevetoxin B [11,25–27] and Aflatoxins [28,29] did not use in ITs construction. Up to now, four generations of ITs produced via four different approaches. The first generation of IT has been developed by attaching the native toxin to full-length mAbs through chemical methods. The ITs had some problems such as low specificity and stability, heterogeneity, reactivity to normal cells, and immunogenicity. Due to these problems, the second generation of IT has been developed. In this generation, the modified toxin, without the natural binding domain, chemically bonded to full-length mAbs. Although the specificity increased, other problems remained [11,30,31]. Third-generation produced by recombinant DNA technology. In this generation, the truncated toxins, without the natural receptor-binding domain, linked to antibody fragments by the peptide linker that led to developing recombinant ITs (RITs) [32,33]. For immunogenicity reduction of RITs, fourth-generation was developed using humanised or fully human formats of antibodies and endogenous proteins of human origin [34,35]. Numerous clinical trials and the US Food and Drug Administration (FDA) approvals indicate the promising IT landscape in cancer treatment (Table 1).
Related Knowledge Centers
- DNA
- DNA Sequencing
- Genome
- Molecular Cloning
- Nucleotide
- Genetic Recombination
- Palindromic Sequence
- Sticky & Blunt Ends
- Species
- Oligonucleotide Synthesis