Expression of genes introduced into cells by retroviral infection is more efficient than that of genes introduced into cells by DNA transfection

Author:

Hwang L H,Gilboa E

Abstract

Calcium phosphate-mediated DNA transfection and retroviral infection are two alternative gene transfer techniques designed to introduce specific DNA fragments into the chromosomes of recipient cells. To compare the efficiency of expression of genes introduced into cells by either of these two techniques, a retrovirus-derived vector was constructed from the genome of Moloney murine leukemia virus by replacing the coding sequences of the envelope gene with the bacterial Neor gene derived from Tn5, termed rEnv-Neor. Expression of the hybrid Neor gene in NIH 3T3 cells after DNA transfection or retroviral infection was determined by measuring the steady-state levels of the corresponding cytoplasmic polyadeylated RNA species. Cells containing one copy of the integrated rEnv-Neor DNA introduced into cells by retroviral infection expressed 10- to 50-fold-higher levels of vector-specific RNA compared with cells harboring one copy of the same DNA derived by DNA transfection. Analysis of the integrated rEnv-Neor DNA with the methylation-sensitive restriction enzyme SmaI has shown that DNA integrated after DNA transfection but not after viral infection is partially methylated, predominantly in the 5' long terminal repeat, the region involved in initiation of transcription.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

Reference35 articles.

1. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease digested hybrids;Berk A.;Cell,1977

2. Biological activity of cloned Moloney sarcoma virus DNA: terminal redundant sequences may enhance transformation efficiency;Blair D. G.;Proc. NatI. Acad. Sci. U.S.A.,1980

3. DNA methylation and the regulation of globin gene expression;Busslinger M.;Cell,1983

4. Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentration of formamide;Casev J.;Nucleic Acids Res.,1976

5. Coffin J. 1982. Structure of the retroviral genome p. 261-368. In R. Weiss. N. Teich. H. Varmus. and J. Coffin (ed.). RNA tumor viruses. Cold Spring Harbor Laboratory. Cold Spring Harbor. N.Y.

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