13 research outputs found
Enhanced labeling density and whole-cell 3D dSTORM imaging by repetitive labeling of target proteins
Variation in resource presence, utilization and reproductive success within a population of European Badgers (Meles meles)
Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo
Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing
Manipulation of proteins by chemical modification is a powerful way to decipher their function. However, most ribosome-dependent and semi-synthetic methods have limitations in the number and type of modifications that can be introduced, especially in live cells. Here, we present an approach to incorporate single or multiple post-translational modifications or non-canonical amino acids into proteins expressed in eukaryotic cells. We insert synthetic peptides into GFP, Na(V)1.5 and P2X2 receptors via tandem protein trans-splicing using two orthogonal split intein pairs and validate our approach by investigating protein function. We anticipate the approach will overcome some drawbacks of existing protein enigineering methods. Chemical modification of proteins can be used to decipher function or use that function for therapeutic purposes. Here, the authors insert synthetic peptides via tandem protein trans-splicing to add post-translational modifications or non-canonical amino acids.Chemical Immunolog