Search PubMed⌕ Search

Biomedical subjects

Akiko Kusunoki

Publications and source records attributed to Akiko Kusunoki.

2 recordsLinked to original sources
PubMed2006

HIV gene therapy using RNA virus systems.

We designed a vector to produce single-stranded DNA (ssDNA). We used HIV-1 reverse transcription for the purpose of constructing a DNAzyme expression vector against the HIV-1 env V3 loop. Initiation of HIV-1 reverse transcription requires the formation of a complex containing the viral RNA, tRNALys and reverse transcriptase. The expression vector contains the HIV-1 primer binding site (PBS) and tRNALys at the 3' end of its RNA transcript, thus enabling to an ssDNA would be synthesized by HIV-1 reverse transcriptase. We have demonstrated that the DNAzyme expressed by the lentiviral vectors suppressed HIV-1 replication in SupT1 cells.

Cell Line↗
PubMed2003

A novel single-stranded DNA enzyme expression system using HIV-1 reverse transcriptase.

In this study, we exploited a DNA enzyme expression system using the mechanism of HIV-1 reverse transcription in vitro. HIV-1 reverse transcription is initiated when its cognate primer tRNA (Lys-3) binds to the primer binding site (PBS) of the viral RNA template. Therefore, this RNA contains the HIV-1 PBS, the DNA enzyme, and a tRNA (Lys-3) at the 3(')-end of its RNA transcript, such that a single-stranded DNA (ssDNA) is synthesized by the HIV-1 reverse transcriptase. We constructed RNA expression vectors including the HIV-1 PBS, the DNA enzyme, and either a native tRNA (Lys-3) or one of two truncated tRNAs (Lys-3), Delta tRNA (Lys-3) and Delta Delta tRNA (Lys-3). The reactions of the pVAX1-Dz-tRNA (Lys-3), pVAX1-Dz-Delta tRNA (Lys-3), and pVAX1-Dz-Delta Delta tRNA (Lys-3) vectors with T7 RNA polymerase in vitro gave the corresponding RNAs. The liberated RNAs were treated with HIV-1 reverse transcriptase (HIV-1 RT) in vitro, which yielded the corresponding ssDNA. The cleavage assay results demonstrated that the expressed DNA enzyme has cleavage ability against the target sequence. Thus, we have found a new DNA enzyme oligonucleotide expression system using the HIV-1 reverse transcriptase in vitro.

Binding Sites↗