A more efficient and specific strategy in the ablation of mRNA in Xenopus laevis using mixtures of antisense oligos.
Previously, antisense oligodeoxyribonucleotides (oligos) have been used to ablate specific mRNAs from the maternal RNA pool of Xenopus laevis oocytes. However, this strategy is limited by the dose of oligo which can be used and the fact that 100% cleavage of the target RNA is rare. Further, non-specific cleavage of other RNAs can also occur. We demonstrate that the use of several oligos against the histone H4 RNA results in a marked improvement in the efficiency of target degradation, due to synergistic action between oligos and the existence of RNA in at least two different secondary structures. We show, by using a set of overlapping oligos complementary to the entire H4 RNA, that the amount of oligo required for efficient target ablation is greatly lowered and non-specific effects are reduced.
Morgan
R
R
School of Biochemistry, University of Birmingham, UK.
Edge
M
M
Colman
A
A
D000818:Animals; D001483:Base Sequence; D005818:Genetic Engineering / Q000379:methods*; D006657:Histones / Q000235:genetics*; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D013329:Structure-Activity Relationship; D014982:Xenopus laevis / Q000235:genetics*
D006657:Histones; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D016914:Ribonuclease H