Detection of Genes Coding for Human Differentiation Markers by Their Transient Expression after DNA Transfer

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 79; no. 1; pp. 146 - 150
• Main Authors: L. J.-A. Chang, Gamble, C. L., Izaguirre, C. A., Minden, M. D., Mak, T. W., McCulloch, E. A.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01.01.1982; National Acad Sciences
• Subjects: Animals; Antibodies, Monoclonal; Blasts; Cell Differentiation; Cell lines; Cells, Cultured; Cellular immunity; Cycloheximide - pharmacology; Dactinomycin - pharmacology; Differentiation antigens; DNA; DNA Restriction Enzymes; DNA, Neoplasm - genetics; Enzymes; Fluorescent Antibody Technique; Kinetics; L cells; Leukemia, Lymphoid - genetics; Leukemia, Myeloid, Acute - genetics; Mice; Monoclonal antibodies; Protein Biosynthesis - drug effects; Spleen cells; T lymphocytes; Transcription, Genetic - drug effects; Transfection
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.79.1.146

We have developed an assay for specific genes in DNA based on transient expression. DNA prepared from patients with acute myeloblastic or acute lymphoblastic leukemia or from the continuous leukemic cell line HL60 was transferred to LTA cells; 48-72 hr later, these recipients expressed hemopoietic differentiation markers as detected by monoclonal antibodies against My-1 (granulocyte specific) and OK-T3 (T-cell specific) using immunofluorescence. The efficiency of transfer was dose and time dependent. We found that genes not expressed in the original cells were expressed after transfer by using this assay. Restriction enzyme analysis showed that My-1 was not expressed with DNA that had been incubated before transfer with either HindIII or Sal I but was present after digestion with either EcoRI or BamHI; after digestion of DNA with the same enzymes, OK-T3 expression was observed only in the HindIII-treated DNA. These studies indicate that DNA was transferred; this approach may provide an efficient method for detecting the activity of specific genes in the absence of selection.