TY - JOUR
T1 - Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: Anti-proliferative effects on HPV 16-transformed cells
AU - Venturini, Francesca
AU - Braspenning, Joris
AU - Homann, Matthias
AU - Gissmann, Lutz
AU - Sczakiel, Georg
N1 - Funding Information:
We thank O. Wildner for initial support as well as A. Hörster and R. Kronenwett for help with FACS analysis and transfection experiments. We also thank B. Teichmann and V. Patzel for critical comments on the manuscript. This work was supported by the Bundesministerium für Bildung, Forschung und Wissenschaft (grant no. 01KV9517) and the Fonds der Chemischen Industrie.
PY - 1999/4/1
Y1 - 1999/4/1
N2 - The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 104 M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.
AB - The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 104 M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.
UR - http://www.scopus.com/inward/record.url?scp=0033118276&partnerID=8YFLogxK
U2 - 10.1093/nar/27.7.1585
DO - 10.1093/nar/27.7.1585
M3 - Journal articles
C2 - 10075988
AN - SCOPUS:0033118276
SN - 0305-1048
VL - 27
SP - 1585
EP - 1592
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 7
ER -