Hairpin Antisense Oligonucleotides Containing 2′-Methoxynucleosides with Base-Pairing in the Stem Region at the 3′-end: Penetration, Localization, and Anti-HIV Activity

Kuwasaki, Tomoyuki; Hosono, Kuniaki; Takai, Kazuyuki; Ushijima, Kaoru; Nakashima, Hideki; Saito, Takeshi; Yamamoto, Naoki; Takaku, Hiroshi

doi:10.1006/bbrc.1996.1707

Cited by 16 publications

(7 citation statements)

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“…In this study, we used a long dsRNA, in place of antisense DNA (31,32), to interfere with the expression of HIV-1 genes. We found that the greatest inhibitory effects were detected with 531 bp E2-dsRNA containing the major CD4-binding domain sequence of gp120, as the target of the HIV-1 env gene.…”

Section: Introductionmentioning

confidence: 99%

Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference

Park¹

2002

Nucleic Acids Research

113

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The RNA interference (RNAi) phenomenon is a recently observed process in which the introduction of a double-stranded RNA (dsRNA) into a cell causes the specific degradation of a mRNA containing the same sequence. The 21-23 nt guide RNAs, generated by RNase III cleavage from longer dsRNAs, are associated with sequence-specific mRNA degradation. Here, we show that dsRNA specifically suppresses the expression of HIV-1 genes. To study dsRNA-mediated gene interference in HIV-1-infected cells, we have designed six long dsRNAs containing the HIV-1 gag and env genes. HIV-1 replication was totally suppressed in a sequence-specific manner by the dsRNAs in HIV-1-infected cells. Especially, E2 dsRNA containing the major CD4-binding domain sequence of gp120, as the target of the HIV-1 env gene, dramatically inhibited the expression of the HIV-1 p24 antigen in PBMCs for a relatively long time. The dsRNA interference method seems to be a promising new strategy for anti-HIV-1 gene therapeutics.

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Section: Introductionmentioning

confidence: 99%

Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference

Park¹

2002

Nucleic Acids Research

113

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“…This activity was strong compared with oligonucleotides targeted to the splice acceptor of the tat gene and the AUG initiation codon of the rev gene (38). We have also shown in vitro that mammalian tRNase ZL with the aid of a 5′-half-tRNA-like sgRNA can cleave a partial HIV-1 mRNA substrate containing the p24 site of the HIV-1 gag gene (24).…”

Section: Resultsmentioning

confidence: 99%

Inhibition of HIV-1 gene expression by retroviral vector-mediated small-guide RNAs that direct specific RNA cleavage by tRNase ZL

Habu

Miyano-Kurosaki²,

Kitano³

et al. 2005

Nucleic Acids Research

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The tRNA 3′-processing endoribonuclease (tRNase Z or 3′ tRNase; EC 3.1.26.11) is an essential enzyme that removes the 3′ trailer from pre-tRNA. The long form (tRNase ZL) can cleave a target RNA in vitro at the site directed by an appropriate small-guide RNA (sgRNA). Here, we investigated whether this sgRNA/tRNase ZL strategy could be applied to gene therapy for AIDS. We tested the ability of four sgRNA-expression plasmids to inhibit HIV-1 gene expression in COS cells, using a transient-expression assay. The three sgRNAs guide inhibition of HIV-1 gene expression in cultured COS cells. Analysis of the HIV-1 mRNA levels suggested that sgRNA directed the tRNase ZL to mediate the degradation of target RNA. The observation that sgRNA was localized primarily in nuclei suggests that tRNase ZL cleaves the HIV-1 mRNA when complexed with sgRNA in this location. We also examined the ability of two retroviral vectors expressing sgRNA to suppress HIV-1 expression in HIV-1-infected Jurkat T cells. sgRNA-SL4 suppressed HIV-1 expression almost completely in infected cells for up to 18 days. These results suggest that the sgRNA/tRNase ZL approach is effective in downregulating HIV-1 gene expression.

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“…In previous studies, the random mixtures of diastereomers S‐ODNs complementary to the gag mRNA containing the AUG initiation codon sequence were more effective inhibitors of HIV‐1 replication than those targeted to the splice‐acceptor site of the tat gene and the AUG initiation codon of the rev gene in acutely infected cells [32]. Thus, we evaluated the anti‐HIV‐1 activity by the inhibition of virus‐specific antigen expression in long‐term HIV‐1 IIIB ‐infected MOLT‐4 clone 8 cells treated with three different target oligonucleotides ([Mix]‐S‐ODN‐ gag ‐28‐AUG‐as, [Mix]‐S‐ODN‐ tat ‐28‐sa‐as, and [Mix]‐S‐ODN‐ rev ‐28‐AUG‐as).…”

Section: Resultsmentioning

confidence: 99%

“…In particular, we first succeeded in the treatment of influenza in an experimental infectious mouse model of influenza A virus, using S‐ODNs [30,31]. We also demonstrated their effectiveness as inhibitors of HIV‐1 replication in an acute‐infection assay [24,32].…”

Section: Introductionmentioning

confidence: 98%

Inhibition of human immunodeficiency virus type 1 replication by P‐stereodefined oligo(nucleoside phosphorothioate)s in a long‐term infection model

et al. 2002

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Oligo(nucleoside phosphorothioate)s (S-ODNs), if prepared by conventional methods, consist of a mixture of diastereomers by virtue of the asymmetry of the phosphorus atom involved in the internucleotide linkages. This may affect the stability of the complexes formed between S-ODNs and complementary oligoribonucleotides, which is commonly accepted as the most important factor in determining the efficacy of an antisense approach. Using HIV-1-infected MOLT-4 cells via a long-term culture approach, we studied the influence of the P-chirality sense of stereodefined 28mer oligo(nucleoside phosphorothioate)s, [All-Rp]-S-ODN-gag-28-AUG and [All-Sp]-S-ODN-gag-28-AUG, complementary to the sequence starting at the AUG initiation codon of the gag mRNA of HIV-1, upon the anti-HIV-1 activity. The [All-Sp]-S-ODN-gag-28-AUG at a low concentration of 0.5 microM can completely suppress HIV-1(gag) p24 antigen expression in HIV-1-infected MOLT-4 clone 8 cells for 32 days. Cells treated with [All-Rp]-S-ODN-gag-28-AUG (0.5 microM) showed a high level of the antigen expression at day 16. Furthermore, satisfactory suppression could not be achieved from a random [Mix]-S-ODN-gag-28-AUG, consisting of a diastereomeric mixture of the oligonucleotides. Our results suggest that chemotherapy based upon the use of stereodefined antisense [All-Sp] S-ODN may be a more effective method for reducing the viral burden in HIV-1-infected individuals.

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Hairpin Antisense Oligonucleotides Containing 2′-Methoxynucleosides with Base-Pairing in the Stem Region at the 3′-end: Penetration, Localization, and Anti-HIV Activity

Cited by 16 publications

References 0 publications

Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference

Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference

Inhibition of HIV-1 gene expression by retroviral vector-mediated small-guide RNAs that direct specific RNA cleavage by tRNase ZL

Inhibition of human immunodeficiency virus type 1 replication by P‐stereodefined oligo(nucleoside phosphorothioate)s in a long‐term infection model

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