Plants Express Anti AGR2 Antibody 18A4 And CRISPR/CAS9 Knock Out XylT And FucT Gene Of Carrot Genomic DNA

Researches revealed that AGR2 protein is overexpressed in most glandular cancers,and related to many cancer symptoms.Articles reported that AGR2 is able to indicate early breast cancer,and becomes one of the most potential target proteins in many cancers.Our lab has already constructed the plasmid which can be transfected in CHO cells to produce humanized AGR2 antibody 18A4.However,it takes much time and money to use animal cells to produce antibody which also has a low yield.FDA approved first recombinant pharmaceutical protein Elelyso which is produced by carrot suspension cells in large scale,to treat Gaucher’s disease(GD).Our project aims to construct a plasmid which can be transfected in plants to express 18A4 and optimize the structure of the plasmid.The results show that the plasmid constructed has 18A4 expression that can be successfully detected by western blotting.Meanwhile,expression of optimized plasmid has apparently increases.However,structure of plants glycosylation still differs from protein from human.Plants have extra structures of Xylose and Fucose.Our research also aims to utilize CRISPR/Cas9 technology to knock out transferase gene of Xylose and Fucose in genomes of carrot suspension cells in purpose to avoid reverse reactions caused by difference of glycosylation.Our results have great significance to express pharmaceutical protein in plants and construct Xyl T and FucT gene knock-out carrot suspension cells.