{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Blumberg LC"],"funding":["Hudson River Biotechnology B.V.","HORIZON EUROPE Framework Programme","Horizon Europe","Dutch Research Council (NWO)"],"pagination":["239-255"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12854893"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["24(1)"],"pubmed_abstract":["Nicotiana benthamiana serves as a unique platform for biopharmaceutical production, offering advantages such as efficient and scalable protein synthesis. In addition, custom N-glycans can be engineered on biopharmaceutical glycoproteins. Yet, plant-native glycosyltransferases and glycoside hydrolases need to be removed to prevent undesired modifications of tailored N-glycans. CRISPR-based systems offer tremendous potential; however, the ploidy of the allotetraploid N. benthamiana can make genome editing challenging when attempting to knock out multiple undesired enzymes using transgenes. Here, we report a highly efficient CRISPR ribonucleoprotein (RNP)-protoplast genome editing strategy for rapid, single-generation platform engineering. We delineate the editing characteristics of ErCas12a "],"journal":["Plant biotechnology journal"],"pubmed_title":["Highly efficient transgene-free ErCas12a RNP-protoplast genome editing and single-cell regeneration in Nicotiana benthamiana for glyco-engineering."],"pmcid":["PMC12854893"],"funding_grant_id":["16740","101080784"],"pubmed_authors":["Bakker GM","Gariepy E","van de Geest H","Wilbers RHP","Slootweg EJ","Los FCO","Blumberg LC","Westerhof LB","van der Kaaij A"],"additional_accession":[]},"is_claimable":false,"name":"Highly efficient transgene-free ErCas12a RNP-protoplast genome editing and single-cell regeneration in Nicotiana benthamiana for glyco-engineering.","description":"Nicotiana benthamiana serves as a unique platform for biopharmaceutical production, offering advantages such as efficient and scalable protein synthesis. In addition, custom N-glycans can be engineered on biopharmaceutical glycoproteins. Yet, plant-native glycosyltransferases and glycoside hydrolases need to be removed to prevent undesired modifications of tailored N-glycans. CRISPR-based systems offer tremendous potential; however, the ploidy of the allotetraploid N. benthamiana can make genome editing challenging when attempting to knock out multiple undesired enzymes using transgenes. Here, we report a highly efficient CRISPR ribonucleoprotein (RNP)-protoplast genome editing strategy for rapid, single-generation platform engineering. We delineate the editing characteristics of ErCas12a ","dates":{"release":"2026-01-01T00:00:00Z","publication":"2026 Jan","modification":"2026-06-17T07:00:19.773Z","creation":"2026-06-17T03:10:24.195Z"},"accession":"S-EPMC12854893","cross_references":{"pubmed":["40522814"],"doi":["10.1111/pbi.70141"]}}