Research Group: Protein O-glycosylation in Plants

• Gene discovery & substrate specificity characterisation of extensins and ArabionoGalactan O-glycosylation enzymes
• Implementation of ‘open ended discovery strategies’ for mining of the plant O-glycome as outlined by our collaborator Copenhagen Center for Glycomics (http://glycomics.ku.dk)
• Implementation and optimization of precise genetic engineering technologies in plants
• Glycoengineering tobacco for production of therapeutic glycoproteins with compatible or defined immune stimulatory glycans
• Gene editing of Danish potato elite cultivars for increased resistance to late blight (Phytoftora infestans) and designer starch qualities

• GUDP
• NNF

One arm is our gene editing effort of Danish potato elite cultivars for increased resistance to late blight (Phytoftora infestans) and designer starch qualities – as outlined in the GUDP and NNF grants described above.

Our long standing glycogene discovery effort has continuously resulted in identification of new glycogenes that control extensin type glycosylation.

As another arm of our translational efforts, we have spearheaded engineering of mammalian type O-glycosylation in plants with the aim of producing a cancer vaccine against the immunogenic cancer specific Tn glycoprotein epitope. Initiation of mammalian type O-glycosylation was engineered transiently in Australian tobacco and stably in tobacco suspension cells and Arabidopsis as evidenced by O-glycosylation of a number of therapeutically relevant substrates. Extensin type glycosylation was also identified on the therapeutic glycoproteins, and we have shown that these glyoepitopes, which are considered adverse/allergenic to man, are prevalent on a number of major plant pollen allergens.

As a yet another arm of our translational efforts we have, together with CCG and world leading allergy company Alk-abello, used engineered defined glycosylation on major protein allergens to target lectin receptors on Antigen Presenting Cells (APCs) of the innate immune system with the downstream aim of improving efficacy and providing novel therapeutic properties.

• Larsen JS, Karlsson RTG, Weihua Tian, Schulz MA, Matthes A, Clausen H, Yang Z, Petersen BL (2019) Engineering Mammalian Cells to Produce Plant-Specific N-glycosylation on Proteins. Glycobiology. In press

• Mnich E, Bjarnholt N, Eudes A, Harholt J, Holland C, Jørgensen B, Larsen FH, Liu M, Manat R, Meyer AS, Mikkelsen JD, Motawie MS, Muschiol J, Møller BL, Møller SR, Perzon A, Petersen BL, Ravn JL, Ulvskov P (2019) Phenolic cross-links: Building and de-construction of the plant cell wall. Natural Product Reports, 2020, DOI: 10.1039/C9NP00028C

• Bennett EP, Petersen BL, Johansen IE, Zhang Y, Chamberlain CA, Özcan Met, Wandall HH, Frodin M (2020) INDEL detection, the “Achilles heel” of precise genome editing: A survey of the available methods for accurate profiling of indels in mono- to polyploid cells. Nucleic Acid Res. (invited review). Provisionally accepted

• Johansen IE, Liu Y, Jørgensen B, Nielsen KL, Andreasson E, Bennett EP, Nielsen Blennow A, Petersen BL (2019) High efficacy full allelic CRISPR/Cas9 gene editing in tetraploid potato (2019) Sci Rep 9, 17715 https://doi.org/10.1038/s41598-019-54126-w 

• Petersen BL, Möller SR, Mravec J, Jørgensen B, Christensen M, Liu Y, Wandall HH, Bennett EP, Yang, Z (2019) Improved CRISPR/Cas9 gene editing by fluorescence activated cell sorting of green fluorescence protein tagged protoplasts. BMC Biotechnology 19:36, ttps://doi.org/10.1186/s12896-019-0530-x.

• Mathiesen CBK, Carlsson MC, Brand S, Möller SR, Idorn M, Straten PT, Pedersen AE, Dabelsteen S, Halim A, Wurtzen PA, Brimnes J, Ipsen H, Petersen BL, Wandall HH (2018) Genetically engineered cell factories produce glycoengineered vaccines that target antigen presenting cells and alter immunity. J Allergy and Clin Immunol. 1016/j.jaci.2018.07.030

• Moeller SR, Yi X, Velásquez SM, Gille S, Hansen PLM, Poulsen C, Olsen CE, Rejzek M, Parsons H, Zhang Y, Wandall HH, Clausen H, Field RA, Pauly M, Estevez JM, Harholt J, Ulvskov P, Petersen BL. Identification and evolution of a plant cell wall specific glycoprotein glycosyl transferase, ExAD. 2017 Sci. Rep, doi: http://dx.doi.org/10.1038/srep45341

• Halim A, Larsen ISB, Neubert P, Joshi H., Petersen BL, Vakhrushev SY, Strahl S, Clausen H (2015) Discovery of a Nucleocytoplasmic O-Mannose Glycoproteome in Yeast. Proc Natl Acad Sci USA 112 (51): 15648–15653

• Halim A, Carlsson MC, Madsen CB, Brand S, Møller SR, Olsen CE, Vakhrushev SY, Clausen H, Brimnes J, Wurtzen PA, Ipsen H, Wandall H, Petersen BL (2015) Top-down and bottom-up mass spectrometry of seven major allergenic proteins reveal novel post translational modifications. Mol Cell Proteomics 14:191-204