Inhibition of vesicular stomatitis virus in cells constitutively expressing an antisense RNA targeted against the virus RNA polymerase gene.

Takacs, Adrienne M.; Banerjee, Amiya K.

doi:10.1099/0022-1317-78-1-125

Cited by 4 publications

(2 citation statements)

References 19 publications

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“…This hypothesis is supported by the recent finding of significant inhibition of vesicular stomatitis virus (VSV) propagation by antisense L-gene sequences (38). Therefore, the presence of the mutated N/P gene border directing efficient transcriptional stop but no detectable restart might be essential for efficient propagation of SAD Ambi-CAT.…”

Section: Fig 5 Quantitation Of Genome and Antigenome Rnas In Virus-mentioning

confidence: 86%

Ambisense gene expression from recombinant rabies virus: random packaging of positive- and negative-strand ribonucleoprotein complexes into rabies virions

Finke¹,

Conzelmann²

1997

J Virol

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We have recovered from cDNA a rabies virus (RV) containing identical, transcriptionally active promoters at its genome (negative-strand) and antigenome RNA and directing efficient expression of a chloramphenicol acetyltransferase (CAT) reporter gene from the antigenome. Transcription of the antigenome CAT gene was terminated by a modified RV gene junction able to mediate transcription stop and polyadenylation but not reinitiation of downstream transcripts. While in standard RV-infected cells genome and antigenome RNAs were present in a 49:1 ratio, the ambisense virus directed synthesis of equal amounts of genome and antigenome RNA (1:1). Total replicative synthesis was reduced by a factor of less than 2, revealing an unexpectedly high level of replication activity of the transcriptionally active promoter in the absence of the parental antigenome promoter. Successful packaging of ambisense ribonucleoprotein complexes (RNPs) into virions demonstrated that the parental 5 end of the RV genome RNA does not contain putative signals required for incorporation into virions. As determined both for standard RV and ambisense RV, virus particles contained genome and antigenome RNPs in the same ratios as those present in infected cells (49:1 and 1:1, respectively), indicating indiscriminate incorporation of RNPs independent of signals in the RNA. Ambisense expression of multiple foreign genes from RV vectors may circumvent problems with transcriptional attenuation of rhabdovirus housekeeping genes.

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Section: Fig 5 Quantitation Of Genome and Antigenome Rnas In Virus-mentioning

confidence: 86%

Ambisense gene expression from recombinant rabies virus: random packaging of positive- and negative-strand ribonucleoprotein complexes into rabies virions

Finke¹,

Conzelmann²

1997

J Virol

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“…The use of a relatively higher oligonucleotide concentration for VSV in the present study may be related to the intrinsic high growth rate of this virus. In a recent study, Tackas and Banerjee (45) could demonstrate the inhibition of VSV in cells constitutively expressing an antisense RNA targeted against the virus L protein gene only at a very low MOI of 0.01 to 0.1 and concluded that the "robust growth rate of VSV eventually overwhelms the available antisense RNA and leads to delayed cell death." FIG.…”

Section: Discussionmentioning

confidence: 99%

Oligonucleotides Tethered to a Short Polyguanylic Acid Stretch Are Targeted to Macrophages: Enhanced Antiviral Activity of a Vesicular Stomatitis Virus-Specific Antisense Oligonucleotide

Prasad

Hashim

Mukhopadhyay

et al. 1999

Antimicrob Agents Chemother

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The poor membrane permeability of oligonucleotides is one of the major problems of antisense technology. Here we report the construction of designer oligonucleotides for targeted delivery to macrophages. The oligonucleotides tethered to a 10-mer poly(G) sequence at their 3′ ends were recognized by scavenger receptors on macrophages and were taken up about 8- to 10-fold as efficiently as those oligonucleotides that either lacked a poly(G) tail or that contained a 10-mer poly(C) tail instead of the poly(G) tail. The enhanced uptake of poly(G) constructs was inhibited in the presence of poly(G) and other known ligands of the scavenger receptor. The bioefficacy of poly(G)-mediated targeting of antisense oligonucleotides (ANS) was demonstrated by using vesicular stomatitis virus (VSV) as a model system. The ability of ANS directed against the translation initiation site of N protein mRNA of VSV to inhibit virus replication was assessed. The ANS with the 10-mer poly(G) sequences (ANS-G) brought about significant inhibition of VSV replication in J774E cells (a murine monocyte/macrophage cell line) and Chinese hamster ovary (CHO) cell transfectants expressing scavenger receptors. The ANS lacking a 10-mer poly(G) stretch were ineffective. The inhibition of VSV replication due to ANS-G was completely abrogated in the presence of 10-mer poly(G), indicating that the antisense effect of the ANS-G molecule was a consequence of scavenger receptor-mediated enhanced uptake. Importantly, antisense molecules linked exclusively by natural phosphodiester bonds were as bioeffective as those synthesized with a mixed backbone of phosphodiester and phosphorothioate. Taken together, these results suggest that macrophage-directed designer ANS against infective agents may simply be obtained by adding a short stretch of guanylic acid sequence to the desired specific ANS during solid-phase synthesis. This nucleic acid-based strategy, which utilizes homogeneous preparation of ANS, may find applications in directed manipulation of macrophage metabolism for a variety of purposes as well as in therapy of a broad spectrum of macrophage-related disorders amenable to the antisense approach.

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Rhabdoviruses

Pringle¹

2003

Encyclopedia of Life Sciences

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The viruses classified in the family Rhabdoviridae have monopartite, negative‐sense, single‐stranded ribonucleic acid (RNA) genomes, 11–15 kb in size; the virions have helical nucleocapsids contained within bullet‐shaped or bacilliform enveloped particles. Rhabdoviruses infect animals, plants and invertebrates; some rhabdoviruses multiply in both invertebrates and vertebrates, and others in both invertebrates and plants, but none multiply in all three hosts. The family includes several viruses responsible for considerable economic losses in agriculture and commercial fisheries; Rabies virus is the most significant human pathogen.

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Inhibition of vesicular stomatitis virus in cells constitutively expressing an antisense RNA targeted against the virus RNA polymerase gene.

Cited by 4 publications

References 19 publications

Ambisense gene expression from recombinant rabies virus: random packaging of positive- and negative-strand ribonucleoprotein complexes into rabies virions

Ambisense gene expression from recombinant rabies virus: random packaging of positive- and negative-strand ribonucleoprotein complexes into rabies virions

Oligonucleotides Tethered to a Short Polyguanylic Acid Stretch Are Targeted to Macrophages: Enhanced Antiviral Activity of a Vesicular Stomatitis Virus-Specific Antisense Oligonucleotide

Rhabdoviruses

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