Enhancement or inhibition of HIV-1 replication by intracellular expression of sense or antisense RNA targeted at different intermediates of reverse transcription

Peng, Hairong; Callison, Deborah; Burrell, Christopher J.

doi:10.1097/00002030-199705000-00006

Cited by 13 publications

(2 citation statements)

References 35 publications

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“…tRNA promoters are pol III type 2 promoters and have widely been used to express ribozymes and antisense RNA [88,[109][110][111][112]. They produce longer transcripts (around 100 nt) compared to type 3 shRNA.…”

Section: Trna Promotersmentioning

confidence: 99%

In Search of the Most Suitable Lentiviral shRNA System

Bos

Bruyne

Heirman³

et al. 2009

CGT

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A decade after its discovery, RNA interference has proven to be an instant success both in fundamental research and clinical applications. Lentiviral delivery of shRNAs is one of the most popular approaches to study gene functionalities in both developmental biology and disorders. During the past 10 years, several adaptations and novel techniques have emerged to improve (conditional) transgene expression and to meet researchers' needs. However, due to this magnitude of diversity, it is sometimes difficult to select the most suitable approach for a specific experimental setup. Here, we summarize the different systems and techniques available for every step in the generation of shRNA-bearing lentiviruses. The most crucial point is inevitably the selection of the target sequence itself. A good shRNA design is indispensable and determines almost completely the success of the experiments. In addition, an adequate promoter that drives the shRNA expression has to be chosen depending on its strength, inducibility, tissue-specificity, At this point, the researcher has also to decide whether the expression of the shRNA should be inducible or not. Another point one has to keep in mind is the choice of lentiviral vector in which the silencing cassette will be incorporated; single- or double-copy vectors are available. The last 2 years, shRNA multiplex approaches in which several targets are silenced with one vector have emerged and have shown a lot of potential in complex studies (like HIV-1). Finally, in the last section, we will discuss the possible induction of an immune response by short dsRNA molecules.

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Section: Trna Promotersmentioning

confidence: 99%

In Search of the Most Suitable Lentiviral shRNA System

Bos

Bruyne

Heirman³

et al. 2009

CGT

View full text Add to dashboard Cite

show abstract

“…Met -antisense RNAs targeted against the HIV-1 U3-R-U5 (684-nts) and U3-R-U5-Psi (800-nts) regions were also developed (135). Stable transductants expressing either one of the two antisense RNAs were shown to inhibit virus replication for 20 weeks post-infection (135).…”

Section: Double Copy Retroviral Vectors Expressing Trnaimentioning

confidence: 99%

Current develoments and future prospects for HIV gene therapy using interfering RNA-based strategies

Joshi¹

2000

Front Biosci

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Effect of deletion and the site of insertion in double copy anti-tat retroviral vectors: viral titres and production of anti-tat mRNA

Carr

Calvert

Kumar

et al. 2001

Archives of Virology

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In attempts to further develop murine leukemia virus (MLV) based retroviral vectors for gene therapy, we investigated vector production and antisense expression from retroviral constructs with U3 deletions or insertions. Promoter elements in the U3 region of the 3' LTR of the vector pLXSN were deleted and replaced with DNA encoding the HIV anti-tat gene under control of the tRNAmet promoter to produce a double copy self inactivating vector (DC-SIN). DC-SIN constructs were compared to vectors containing the anti-tat cassette inserted at 5 different sites of the U3 region (DC-insertions). Titres of DC-SIN and DC-insertion vectors were similar but approximately 10 fold lower than parental pLXSN. Cells transduced with DC-SIN and DC-insertion vectors all expressed anti-tat mRNA. Transcripts from the MLV-LTR were detected in cells transduced with DC-insertion but not DC-SIN vectors or a vector with the anti-tat cassette between CAAT and TATA boxes of the promoter, indicating inactivation of the viral promoter in the latter vectors. Cells transduced with constructs of either design showed comparable efficacy of protection against HIV challenge. Thus, no U3 insertion site was preferred for virus production. Insertion of a tRNA promoter between CAAT and TATA boxes and the DC-SIN design which would not introduce an active RNA pol II promoter into the genome are attractive for further development of safe gene therapy agents.

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Enhancement or inhibition of HIV-1 replication by intracellular expression of sense or antisense RNA targeted at different intermediates of reverse transcription

Abstract: Plus-strand strong-stop DNA as an intermediate in the early process of viral reverse transcription can be explored as an additional target for anti-HIV gene therapy.

Cited by 13 publications

References 35 publications

In Search of the Most Suitable Lentiviral shRNA System

In Search of the Most Suitable Lentiviral shRNA System

Current develoments and future prospects for HIV gene therapy using interfering RNA-based strategies

Effect of deletion and the site of insertion in double copy anti-tat retroviral vectors: viral titres and production of anti-tat mRNA

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