Johnson L, McPhee D, Purcell D; Australasian Society for HIV Medicine. Conference.
Annu Conf Australas Soc HIV Med. 1996 Nov 14-17; 8: 116 (poster no. 133).
Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria.
Human immunodeficiency virus type-1 (HIV-1) RNA undergoes a variety of post-transcriptional processing events, each of which represents a potential intervention point for antiviral therapy. Antisense oligonucleotides, ribozymes and antisense messenger RNAs have all been used to inhibit HIV-1 replication in vitro. Selection techniques have been used in developing these antiviral agents that involve mutagenesis and selection of the most effective antiviral molecules. We aimed to extend the generation of high affinity oligonucleotides and oligoribonucleotides by in vitro selection. We have developed a technique for generating oligoribonucleotide pools of random sequence and length that bind to HIV-1 RNA. Oligonucleotides were designed with fixed 5' ends that include either a T7 or SP6 RNA polymerase binding site and fixed 3' ends with intervening random sequences. These oligonucleotides were used to self anneal in a polymerase chain reaction (PCR) to generate random, double-stranded oligonucleotides of variable or fixed length, depending on conditions used. Oligonucleotides or transcribed oligoribonucleotides were mixed with HIV-1 RNA, separated by electrophoresis, and the HIV-1 binding nucleotide sequences recovered by a reverse-transcriptase PCR reaction. The technique allows identification and characterization of the most exposed regions of RNA. It is therefore useful as an analytical tool for studying HIV-1 RNA secondary structure along with identifying new targets for antiviral therapy.
Publication Types:
Keywords:
- Base Sequence
- DNA-Directed RNA Polymerases
- HIV-1
- Humans
- In Vitro
- Oligonucleotides
- Oligonucleotides, Antisense
- Oligoribonucleotides
- Polymerase Chain Reaction
- RNA
- RNA Processing, Post-Transcriptional
- RNA polymerase SP6
- RNA, Antisense
- RNA, Catalytic
- RNA, Complementary
- RNA, Messenger
- RNA-Directed DNA Polymerase
- Selection (Genetics)
- genetics
- reverse transcriptase, Human immunodeficiency virus 1
Other ID:
UI: 102221971
From Meeting Abstracts