Nucleic Acids Research, Vol 24, Issue 21 4117-4122, Copyright © 1996 by Oxford University Press
GD Jones, EA Lesnik, SR Owens, LM Risen and RT Walker
The thermal stabilities of the duplexes formed between 4'-thio-modified
oligodeoxynucleotides and their DNA and RNA complementary strands were
determined and compared with those of the corresponding unmodified
oligodeoxynucleotides. A 16mer oligodeoxynucleotide containing 10
contiguous 4'-thiothymidylate modifications formed a less stable duplex
with the DNA target (deltaTm/modification -1.0 degrees C) than the
corresponding unmodified oligodeoxynucleotide. However, when the same
oligodeoxynucleotide was bound to the corresponding RNA target, a small
increase in Tm was observed (deltaTm/modification +0.16 degrees C) when
compared with the unmodified duplex. A study to identify the specificity of
an oligodeoxynucleotide containing a 4'-thiothymidylate modification when
forming a duplex with DNA or RNA containing a single mismatch opposite the
modification found the resulting Tms to be almost identical to the
wild-type duplexes, demonstrating that the 4'-thio- modification in
oligodeoxynucleotides has no deleterious effect on specificity. The
nuclease stability of 4'-thio-modified oligodeoxynucleotides was examined
using snake venom phosphodiesterase (SVPD) and nuclease S1. No significant
resistance to degradation by the exonuclease SVPD was observed when
compared with the corresponding unmodified oligodeoxynucleotide. However,
4'-thio-modified oligodeoxynucleotides were found to be highly resistant to
degradation by the endonuclease S1. It was also demonstrated that
4'-thio-modified oligodeoxynucleotides elicit Escherichia coli RNase H
hydrolysis of the RNA target only at high enzyme concentration.
ARTICLES
Investigation of some properties of oligodeoxynucleotides containing 4'- thio-2'-deoxynucleotides: duplex hybridization and nuclease sensitivity
The School of Chemistry, The University of Birmingham, UK.
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