Journal of Microbiology

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Adenoviral Vector System: A Comprehensive Overview of Constructions, Therapeutic Applications and Host Responses: Anyeseu Park, Jeong Yoon Lee; J. Microbiol. 2024;62(7):491-509. Published online July 22, 2024; DOI: https://doi.org/10.1007/s12275-024-00159-4

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Adenoviral vectors are crucial for gene therapy and vaccine development, offering a platform for gene delivery into host cells. Since the discovery of adenoviruses, first-generation vectors with limited capacity have evolved to third-generation vectors flacking viral coding sequences, balancing safety and gene-carrying capacity. The applications of adenoviral vectors for gene therapy and anti-viral treatments have expanded through the use of in vitro ligation and homologous recombination, along with gene editing advancements such as CRISPR-Cas9. Current research aims to maintain the efficacy and safety of adenoviral vectors by addressing challenges such as pre-existing immunity against adenoviral vectors and developing new adenoviral vectors from rare adenovirus types and non-human species. In summary, adenoviral vectors have great potential in gene therapy and vaccine development. Through continuous research and technological advancements, these vectors are expected to lead to the development of safer and more effective treatments.

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Journal Article

Comparative Analysis of Intracellular Trans-Splicing Ribozyme Activity Against Hepatitis C Virus Internal Ribosome Entry Site: Kyung-Ju Ryu Seong-Wook Lee; J. Microbiol. 2004;42(4):361-364.; DOI: https://doi.org/2097 [pii]

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Abstract: Internal ribosome entry site (IRES) of the hepatitis C virus (HCV) is known to be essential for HCV replication and most conserved among HCV variants. Hence, IRES RNA is a good therapeutic target for RNA-based inhibitors, such as ribozymes. We previously proposed a new anti-HCV modulation strategy based on trans-splicing ribozymes, which can selectively replace HCV transcripts with a new RNA that exerts anti-HCV activity. To explore this procedure, sites which are accessible to ribozymes in HCV IRES were previously determined by employing an RNA mapping method in vitro. In this study, we evaluate the intracellular accessibility of the ribozymes by comparing the trans-splicing activities in cells of several ribozymes targeting different sites of the HCV IRES RNA. We assessed the intracellular activities of the ribozymes by monitoring their target-specific induction degree of both reporter gene activity and cytotoxin expression. The ribozyme capable of targeting the most accessible site identified by the mapping studies then harbored the most active trans-splicing activity in cells. These results suggest that the target sites predicted to be accessible are truly the most accessible in the cells, and thus, could be applied to the development of various RNA-based anti-HCV therapies.

Studies on the Inhibition of HIV Replication with a Number of RRE Decoy Derivatives: Lee , Seong Wook; J. Microbiol. 1998;36(4):308-315.

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Abstract: RRE decoys are short RNA oligonucleotides corresponding to the HIV Rev response element (RRE) sequence, which protect cells from HIV replication by inhibiting the binding of the HIV regulatory protein Rev to the authentic HIV RRE region. Previously minimal RRE decoy containing the 13-nucleotide primary Rev binding domain of RRE was described to be a potent inhibitor of HIV in CEM cells. In this report, we analyzed and compared the ability of a series of RRE decoy derivatioves to inhibit HIV replication in CEM cells to develop increasingly effective RRE decoy. Using an improved tRNA cassette to express high level of RRE transcripts in cells, we found that a variant form of stem-loop II(SLII) binding domain of wild type RRE termed RRE40 was more potent than any other RRE decoys previously developed or tested here and protected all cells most effectively from HIV. RRE40 was previously selected in vitro which binds to Rev protein 10-fold better than wild type RRE. CEM cells expressing RRE40 decoy RNAE40 decoys inhibit HIV specifically by sequestering Rev binding to the authentic RRE target in HIV RNA and indicated that RRE40 RNA identified by using in vitro binding studies also binds Rev in cells. These obserbations have important implications for experiments involving optimization of clinical application of RNA decoy based gene therapy protocol against HIV.

Improved Inhibition of Human Immunodeficiency Virus Type 1 Replication by Intracellular Co-overexpression of TAR and RRE Decoys in Tandem Array: Seong-Wook Lee; J. Microbiol. 2003;41(4):300-305.

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Abstract: Intracellular expression of RNA decoys, such as TAR or RRE decoy, has been previously shown to protect immune cells from human immunodeficiency virus type 1 (HIV-1) replication by inhibiting the binding of the HIV-1 regulatory protein to the authentic HIV RNA sequence. However, HIV-1 challenge experiments of primary human T cells, which express the RNA decoy, demonstrated that the cells were only transiently protected, and hence, more improved protocols for HIV-1 inhibition with the RNA decoys need to be developed. In this report, in order to develop a more effective RNA decoy, we analyzed and compared the ability of a series of RNA decoy derivatives in inhibiting HIV-1 replication in CEM cells. Using an improved tRNA cassette to express high levels of RNA decoy transcripts in cells, we found that co-expression of both TAR and RRE decoys, in the form of an aligned sequence in a single transcription cassette, much more potently blocked cells from HIV-1 than the expression of only one kind of RNA decoy. This observation will have an important implication for experiments involving optimization of clinical applications in RNA decoy-based gene therapy against HIV-1.

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