Frequency of missense mutations in the coding region of a eukaryotic gene transferred by retroviral vectors.
A relatively high mutation rate is probably a major factor in the evolutionary success of retroviruses, because it generates the genetic diversity that helps them to cope with changes in the environment. When using recombinant retroviruses as vectors for gene transfer and gene therapy, it is important to consider the implications of this biological characteristic. Until now, the mutation rate has been studied by using noneukaryotic genes as reporters. Here we report point mutations in the human glucose-6-phosphate dehydrogenase (hG6PD) gene transferred by Moloney murine leukemia virus-based vectors into murine bone marrow cells and NIH 3T3 murine fibroblasts. After bone marrow transplantation, we observed an hG6PD with abnormal electrophoretic mobility for 2 out of 34 mice. Next, we studied this phenomenon quantitatively and found 1 electrophoretically abnormal hG6PD variant among 93 independently isolated NIH 3T3 clones, from which we estimate a mutation rate of 1.4 x 10(-5) per base pair per replication cycle. Mutations in the transferred gene can thus contribute to the impairment of the effectiveness of retrovirus-mediated gene transfer.
De Angioletti
Maria
M
Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Rovira
Ana
A
Sadelain
Michel
M
Luzzatto
Lucio
L
Notaro
Rosario
R
D016475:3T3 Cells; D000818:Animals; D001854:Bone Marrow Cells; D005057:Eukaryotic Cells; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D006801:Humans; D007942:Leukemia, Experimental / Q000821:virology; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D020125:Mutation, Missense; D012192:Retroviridae Infections / Q000821:virology; D014412:Tumor Virus Infections / Q000821:virology
D005954:Glucosephosphate Dehydrogenase