CRISPR/Cas9-based Antibody Production in Plants

Nowadays demand of antibody production is increased to cure different diseases including diabetes, hepatitis and cancer. For that different types of systems are used for the expression of antibody production. But these were not improved the antibody production. Plant cells have several benefits in comparison with other eukaryotic cells if it is considered as eukaryotic expression system. As compared to the human cell or other microorganisms, the plant cell is safe and decrease the contamination of antibody production. In addition, plants perform proper post-translational modification as a eukaryotic expression system. But recently, transient expression system is used due to the safe and improve the quality and quantity of antibody production. In transient expression system agroinfiltration method are mostly used. The main issue in antibody production is purification. Because in downstream process antibody is degraded due to the physical and chemical stresses. These issues can be solved with the help of CRISPR/Cas9. Plant antibody can be tagged with the help of CRISPR/Cas9. This review encompasses the applications of CRISPR technology for producing plant-based antibodies.

Communication. Vanillyl alcohol oxidases produced in Komagataella phaffii contain a highly stable noncovalently bound anionic FAD semiquinone

Vanillyl alcohol oxidase (VAO) from Penicillium simplicissimum is a covalent flavoprotein that has emerged as a promising biocatalyst for the production of aromatic fine chemicals such as vanillin, coniferyl alcohol and enantiopure 1-(4’-hydroxyphenyl) alcohols. The largescale production of this eukaryotic enzyme in Escherichia coli has remained challenging thus far. For that reason an alternative, eukaryotic expression system, Komagataella phaffii, was tested. Additionally, to produce novel VAO biocatalysts, we screened genomes for VAO homologues. One bacterial and five fungal sequences were selected for expression, using key active site residues as criteria for their selection. Expression of the putative vao genes in K. phaffii was successful, however expression levels were low (1 mg per litre of culture). Surprisingly, all purified enzymes were found to contain a highly stable, non-covalently bound anionic FAD semiquinone that could not be reduced by dithionite or cyanoborohydride. Activity experiments revealed that VAO expressed in K. phaffii does not produce vanillin because the enzyme suffers from oxidative stress.

Virus-like particles as drug delivery vectors

Virus-like particles (VLPs) assemble spontaneously during the viral cycle or in heterologous systems during expression of viral structural protein. Depending on the complexity of the VLPs, they can be obtained by expression in prokaryotic or eukaryotic expression system from the suitable recombinant vectors, or formed in cell-free conditions. Moreover, they can be built from proteins of a single virus, or can present the proteins or peptides derived from a virus or cell on a platform derived from any other single virus, thus forming chimeric VLPs. VLPs are best known for their immunogenic properties, but the versatility of VLPs allows a wide variety of applications. They are lately in the centre of investigations in vaccinology, drug delivery and gene therapy. This review focuses on utilization of VLPs for drug delivery.

Analysis of Synonymous Codon Usage in the US2 Gene of Duck Plague Virus

The codon usage bias of DPV-CHv US2 gene (GenBank Accession No. EU195086) will be analysed in this study, and a comparative analysis of DPV-CHv US2 gene and those of other 15 alphaherpesvirus US2 genes was performed. We also analysed the RSCU, ENC, GC3s value of those US2 genes in herpesvirus, some data were analysed specifically in the article. Consequently, we found that the eukaryotic expression system——the yeast is more suitable for the expression of DPV US2 gene.