In vitro stability of polymerase chain reaction-generated DNA fragments in serum and cell extracts.
The potential use of polymerase chain reaction (PCR)-generated DNA fragments (PCR-DNAs) as pharmaceutical agents has previously been suggested, with the demonstration of the in vitro cellular internalization and biologic activity of PCR-DNA decoy molecules targeted to human estrogen receptor gene. In order to provide information on the stability of these double-stranded DNA molecules, the nuclease resistance of PCR-DNAs of different sizes was studied in different conditions and experiments. Simulating in vitro and in vivo transfection protocol, we demonstrated that PCR-DNAs exhibited good stability toward fetal bovine serum (FBS) and adult human serum nuclease digestion. In addition, when the protective activity of liposome-based formulations toward nuclease digestion was tested, it was shown that the stability of PCR-DNAs could be further increased (up to 7 days) when a liposome-mediated delivery system was employed.
Piva
R
R
Dipartimento di Biochimica e Biologia Molecolare, Universita di Ferrara, Italy.
Lambertini
E
E
Penolazzi
L
L
Facciolo
M C
MC
Lodi
A
A
Aguiari
G
G
Nastruzzi
C
C
del Senno
L
L
D000328:Adult; D001943:Breast Neoplasms / Q000737:chemistry; D002412:Cations; D002457:Cell Extracts / Q000737:chemistry*; D004247:DNA / Q000032:analysis / Q000097:blood* / Q000737:chemistry*; D004273:DNA, Neoplasm / Q000032:analysis / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry; D015257:DNA-Cytosine Methylases / Q000378:metabolism*; D004355:Drug Stability; D004587:Electrophoresis, Agar Gel; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D008081:Liposomes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012208:Rhabdomyosarcoma / Q000737:chemistry; D014407:Tumor Cells, Cultured
D002412:Cations; D002457:Cell Extracts; D004273:DNA, Neoplasm; D004279:DNA, Viral; D008081:Liposomes; D004247:DNA; D015257:DNA-Cytosine Methylases; C027823:Cytosine 5-methyltransferase