Efficient expression by an alphavirus replicon of a functional ribozyme targeted to human immunodeficiency virus type 1

Smith, Stephen M.; Maldarelli, Frank; Jeang, Kuan‐Teh

doi:10.1128/jvi.71.12.9713-9721.1997

Cited by 17 publications

(3 citation statements)

References 26 publications

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“…Since ribozymes are RNase H-independent, 2′-modifications to increase stability do not diminish antisense effects and experiments have shown some modifications do not attenuate catalytic ability [49]. Unlike AS-ONs, ribozymes can be expressed from a vector, which offers the advantage of continued production of these molecules intracellularly [50,51]. However, stable transformation of cells in vivo has its own complications and will not be discussed in this review.…”

Section: Ribozymesmentioning

confidence: 99%

Suppression of Gene Expression by Targeted Disruption of Messenger RNA: Available Options and Current Strategies

Jen

Gewirtz

2000

STEM CELLS

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At least three different approaches may be used for gene targeting including: A) gene knockout by homologous recombination; B) employment of synthetic oligonucleotides capable of hybridizing with DNA or RNA, and C) use of polyamides and other natural DNA-bonding molecules called lexitropsins.Targeting mRNA is attractive because mRNA is more accessible than the corresponding gene. Three basic strategies have emerged for this purpose, the most familiar being to introduce antisense nucleic acids into a cell in the hopes that they will form Watson-Crick base pairs with the targeted gene's mRNA. Duplexed mRNA cannot be translated, and almost certainly initiates processes which lead to its destruction. The antisense nucleic acid can take the form of RNA expressed from a vector which has been transfected into the cell, or take the form of a DNA or RNA oligonucleotide which can be introduced into cells through a variety of means. DNA and RNA oligonucleotides can be modified for stability as well as engineered to contain inherent cleaving activity.It has also been proven that because RNA and DNA are very similar chemical compounds, DNA molecules with enzymatic activity could also be developed. This assumption proved correct and led to the development of a "general-purpose" RNA-cleaving DNA enzyme. The attraction of DNAzymes over ribozymes is that they are very inexpensive to make and that because they are composed of DNA and not RNA, they are inherently more stable than ribozymes.Although mRNA targeting is impeccable in theory, many additional considerations must be taken into account in applying these strategies in living cells including mRNA site selection, drug delivery and intracellular localization of the antisense agent. Nevertheless, the ongoing revolution in cell and molecular biology, combined with advances in the emerging disciplines of genomics and informatics, has made the concept of nontoxic, cancer-specific therapies more viable then ever and continues to drive interest in this field.

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Section: Ribozymesmentioning

confidence: 99%

Suppression of Gene Expression by Targeted Disruption of Messenger RNA: Available Options and Current Strategies

Jen

Gewirtz

2000

STEM CELLS

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“…Viral genomes engineered for foreign gene expression can be a valuable tool in gene therapy, drug delivery, and vaccine development studies [7][8][9][10]. Viruses expressing heterologous gene(s) of interest can be utilized in high-throughput screening of virus inhibitor compounds or cell-based antiviral effectors and in viral detection tests.…”

Section: Introductionmentioning

confidence: 99%

Novel Approach for Insertion of Heterologous Sequences into Full-Length ZIKV Genome Results in Superior Level of Gene Expression and Insert Stability

Volkova

Tsetsarkin

Sippert

et al. 2020

Viruses

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Zika virus (ZIKV) emerged in the Americas in 2015, presenting unique challenges to public health. Unlike other arboviruses of the Flaviviridae family, it is transmissible by sexual contact, which facilitates the spread of the virus into new geographic areas. Additionally, ZIKV can be transmitted from mother to fetus, causing microcephaly and other severe developmental abnormalities. Reliable and easy-to-work-with clones of ZIKV expressing heterologous genes will significantly facilitate studies aimed at understanding the virus pathogenesis and tissue tropism. Here, we developed and characterized two novel approaches for expression of heterologous genes of interest in the context of full-length ZIKV genome and compared them to two previously published strategies for ZIKV-mediated gene expression. We demonstrated that among the four tested viruses expressing nLuc gene, the virus constructed using a newly developed approach of partial capsid gene duplication (PCGD) attained the highest titer in Vero cells and resulted in the highest level of nLuc expression. Suitability of the PCGD approach for expression of different genes of interest was validated by replacing nLuc sequence with that of eGFP gene. The generated constructs were further characterized in cell culture. Potential applications of ZIKV clones stably expressing heterologous genes include development of detection assays, antivirals, therapeutics, live imaging systems, and vaccines.

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“…SFV expression vectors have been used in a range of mammalian cell lines to express foreign constructs (Smith et al, 1997) (Berglund et al, 1997). To our knowledge this vector system has not been evaluated for use in insect cell lines.…”

Section: Introductionmentioning

confidence: 99%

Semliki Forest virus as an expression vector in insect cell lines

Pettigrew

O'Neill

1999

Insect Molecular Biology

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Studies were undertaken to determine if replication-deficient Semliki Forest virus expression vectors could be successfully used to express foreign gene constructs in insect cell lines. Using green fluorescent protein (GFP) as a marker we recorded infection levels of nearly 100% in the Aedes albopictus cell lines C6/36 and Aa23T, as well as in the Ae. aegypti cell line MOS20. The virus was capable of infecting an Anopheles gambiae cell line MOS55. The amount of GFP protein produced in each cell line was quantified. Northern analysis of viral transcription revealed the presence of novel transcripts in Aa23T, C6/36, and MOS55 cell lines, but not in the BHK or MOS20. The initial characterization of these transcripts is described.

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Efficient expression by an alphavirus replicon of a functional ribozyme targeted to human immunodeficiency virus type 1

Cited by 17 publications

References 26 publications

Suppression of Gene Expression by Targeted Disruption of Messenger RNA: Available Options and Current Strategies

Suppression of Gene Expression by Targeted Disruption of Messenger RNA: Available Options and Current Strategies

Novel Approach for Insertion of Heterologous Sequences into Full-Length ZIKV Genome Results in Superior Level of Gene Expression and Insert Stability

Semliki Forest virus as an expression vector in insect cell lines

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