Potential nuclease-based strategies for HIV gene therapy

Singwi, Sanjeev; Joshi, Sadhna

doi:10.2741/singwi

Cited by 16 publications

(14 citation statements)

References 195 publications

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“…Their application depends on the specificity and mode of action of a particular enzyme. Nucleases have been employed in the production of flavor enhancers, removal of nucleic acids, as therapeutic agents, and for the determination of nucleic acid structure, in particular, mutation typing (Reddy and Shankar 1993;Gite and Shankar 1995;Jasin 1996;Singwi and Joshi 2000;Williams 2001). Nucleases with novel and unusual properties may facilitate the development of advanced technologies in many areas of biotechnology and biomedicine.…”

mentioning

confidence: 99%

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

Shagin¹,

Rebrikov²,

Kozhemyako³

et al. 2002

Genome Res.

229

198

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We have characterized a novel nuclease from the Kamchatka crab, designated duplex-specific nuclease (DSN). DSN displays a strong preference for cleaving double-stranded DNA and DNA in DNA-RNA hybrid duplexes, compared to single-stranded DNA. Moreover, the cleavage rate of short, perfectly matched DNA duplexes by this enzyme is essentially higher than that for nonperfectly matched duplexes of the same length. Thus, DSN differentiates between one-nucleotide variations in DNA. We developed a novel assay for single nucleotide polymorphism (SNP) detection based on this unique property, termed "duplex-specific nuclease preference" (DSNP). In this innovative assay, the DNA region containing the SNP site is amplified and the PCR product mixed with signal probes (FRET-labeled short sequence-specific oligonucleotides) and DSN. During incubation, only perfectly matched duplexes between the DNA template and signal probe are cleaved by DSN to generate sequence-specific fluorescence. The use of FRET-labeled signal probes coupled with the specificity of DSN presents a simple and efficient method for detecting SNPs. We have employed the DSNP assay for the typing of SNPs in methyltetrahydrofolate reductase, prothrombin and p53 genes on homozygous and heterozygous genomic DNA.

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mentioning

confidence: 99%

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

Shagin¹,

Rebrikov²,

Kozhemyako³

et al. 2002

Genome Res.

229

198

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“…71 The specific targeting of proviral DNA in latently infected cells will require genetically engineered nucleases with both a sequence specific binding domain and a catalytically active cleavage domain. While there are a number of highly conserved sequences within the viral genome, including the TAR region 72 and fusion region within Gag, 73 a foremost challenge will be the delivery of these nucleases to viral reservoirs in vivo. Preventing off-target cleavage events is of paramount importance to establish the safety of this approach, yet there are limited options when one considers aggressive measures to neutralize provirus.…”

Section: Targeting Proviral Dna With Genetically Engineered Nucleasesmentioning

confidence: 99%

Genetically Modified Hematopoietic Stem Cell Transplantation for HIV-1–infected Patients: Can We Achieve a Cure?

2014

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The cure of a human immunodeficiency virus (HIV)-1-infected patient following allogeneic transplantation from a CCR5-null donor and potential cure of two patients transplanted with CCR5 wild-type hematopoietic stem cells (HSC) have provided renewed optimism that a potential alternative to conventional antiretroviral therapy (ART) is forthcoming. While allogeneic grafts have thus far suggested complete eradication of viral reservoirs, it has yet to be observed following autologous HSC transplantation. Development of curative autologous transplantation strategies would significantly increase the number of treatable patients, eliminating the need for matched donors and reducing the risks of adverse events. Recent studies suggest gene therapy may provide a mechanism for developing curative therapies. Expression of cellular/artificial restriction factors or disruption of CCR5 has been shown to limit viral replication and provide protection of genetically modified cells. However, significant obstacles remain with regards to the depletion of established viral reservoirs in an autologous transplantation setting devoid of the "allo-effect". Here, we discuss results from early-stage clinical trials and recent findings in animal models of gene modified HSC transplantation. Finally, we propose innovative combination therapies that may aid in the reduction and/or elimination of viral reservoirs in HIV-1-infected patients and promote the artificial development of a natural controller phenotype.

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“…Anti-HIV nucleases act in several ways: they can specifically cleave the HIV-RNA within the cell (targeted nucleases), they can cleave pro-viral DNA or viral progeny RNA by compartmentalizing with the preintegration complex during the early stages of the viral life cycle, or by co-packaging with progeny virus (co-localized nucleases), and finally they can confer selective toxicity to the HIV-infected cells (cytotoxic nucleases). [1] Tev-RNase T1, for example, is a targeted nuclease that can target both TAR and RRE sequences within the HIV-RNA, thus inhibiting the production of both early and late viral gene products. The intracellular production of this nuclease in vitro resulted in a significant delay in replication of HIV-1 without any sign of cellular toxicity [2] (figure 1).…”

Section: Therapeutic Recombinant Genes Expressing Interfering Proteinsmentioning

confidence: 99%

Gene Therapy Approaches to HIV Infection

Friedman

Guallini

Galluzzi

et al. 2002

American Journal of PharmacoGenomics

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The HIV pandemic represents a new challenge to biomedical research. What began as a handful of recognized cases among homosexual men in the US has become a global pandemic of such proportions that it clearly ranks as one of the most destructive viral scourges in history. In the past few years new treatments and drugs have been developed and tested, but the development of a new generation of therapies remains a major priority, because of the lack of chemotherapeutic drugs or vaccines that show long-term efficacy in vivo. Recently, gene therapeutic strategies for the treatment of patients with HIV infection have received increased attention because they are able to offer the possibility of simultaneously targeting multiple sites in the HIV genome, thereby minimizing the production of resistant virus. Recombinant genes for gene therapy can be classified as expressing interfering proteins (intracellular antibodies, dominant negative proteins) or interfering RNAs (antisense RNAs, ribozymes, RNA decoys). The latter group offers the advantage of avoiding the stimulation of host immune response which might progressively decrease the efficacy of proteins. The stumbling block to achieving lasting antiviral effects is still represented by the lack of efficient gene transfer techniques capable of generating persistent transgene expression and a high number of transduced cells relative to untransduced cells. Novel delivery vectors, such as lentiviruses, might overcome some of these shortcomings. The use of recombinant genes to generate immunity is a very promising concept that is rapidly expanding. Since the immune system can significantly amplify the response to tiny amounts of antigen, DNA vaccines can indeed be delivered by exploiting traditional gene therapy approaches without the need of high transduction efficiency.

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Potential nuclease-based strategies for HIV gene therapy

Cited by 16 publications

References 195 publications

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

A Novel Method for SNP Detection Using a New Duplex-Specific Nuclease From Crab Hepatopancreas

Genetically Modified Hematopoietic Stem Cell Transplantation for HIV-1–infected Patients: Can We Achieve a Cure?

Gene Therapy Approaches to HIV Infection

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