Protein microinjection by protease permeabilization of fibroblasts.
Exposure of cultured diploid fibroblasts to protease solutions induces a hyperpermeable state which permits entry of exogenous macromolecules directly into the cytosol. We have exploited this finding to devise a microinjection method whose chief advantages are simplicity and good retention of cell viability. Proteins successfully injected by this technique range from insulin to thyroglobulin. The amounts injected range from 4 X 10(5) to 5 X 10(6) molecules/cell.
Lemons
R
R
Department of Pediatrics, University of Michigan, School of Medicine, Ann Arbor 48109.
Forster
S
S
Thoene
J
J
D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D003012:Microbial Collagenase / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D011402:Pronase / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D011506:Proteins; D010447:Peptide Hydrolases; D014357:Trypsin; D011402:Pronase; D003012:Microbial Collagenase