A SENSITIVE TECHNIQUE TO MONITOR GENE-TRANSFER AND EXPRESSION IN BONE-MARROW STEM-CELLS
- 1 August 1989
- journal article
- research article
- Vol. 17 (7), 832-835
Abstract
The polymerase chain reaction technique (PCR), a primer-mediated enzymatic amplification of specific target sequences, was used to monitor gene transfer into hematopoietic progenitor stem cells. A gene coding for human interleukin 1.alpha. (IL-1.alpha.) was cotransfected with the rous sarcoma virus (RSV) CAT plasmid into mouse bone marrow cells by electroporation. Individual chloramphenicol (CAM)-resistant bone marrow progenitor colonies (granulocyte-macrophage colony-forming units; CFU-GM) containing 50-100 cells were analyzed by PCR for the presence and expression of IL-1.alpha. and CAT sequences. Amplified IL-1.alpha. DNA sequences were detected from a 50-cell CFU-GM colony. CAT and IL-1.alpha. RNA expression was demonstrated from the CAM-resistant CFU-GM colonies.This publication has 10 references indexed in Scilit:
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