Inhibition of HIV-1 replication with designed miRNAs expressed from RNA polymerase II promoters

Hl, Lo; Chang, Tzu-Ming; P, Yam; Pm, Marcovecchio; S, Li; Ja, Zaia; Jk, Yee

doi:10.1038/sj.gt.3303011

Cited by 27 publications

(21 citation statements)

References 38 publications

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“…While many RNAi knockdown vectors designed for general laboratory experimentation use short-hairpin designs, overexpression of shRNAs using RNA polymerase III vectors have been demonstrated to mediate toxicity via saturation of microRNA processing pathways leading to lethality in animal models [35,36]. These shRNA vectors have also been shown to be toxic to transduced human T cells [37] and we also observed this phenomenon in preliminary experiments.…”

Section: Discussionsupporting

confidence: 54%

Gamma-Retroviral Vector Design for the Co-Expression of Artificial microRNAs and Therapeutic Proteins

Park

Abate-Daga²,

Zhang³

et al. 2014

Nucleic Acid Therapeutics

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Section: Discussionsupporting

confidence: 54%

Gamma-Retroviral Vector Design for the Co-Expression of Artificial microRNAs and Therapeutic Proteins

Park

Abate-Daga²,

Zhang³

et al. 2014

Nucleic Acid Therapeutics

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“…Few target genes that are regulated by microRNAs are currently documented, but the ones identified have important roles in apoptosis, homeobox regulation, development [30,[97][98][99][100][101], cell growth and apoptosis [13,14], viral infection [15,19], and human cancer [81,[102][103][104]. This type of gene control represents a new regulatory mechanism and is predicted to affect many crucial cellular processes, including developmental programs.…”

Section: Microrna Regulation Mechanisms and Target Predictionmentioning

confidence: 99%

“…Various types of small, noncoding RNAs exist as modulators of gene expression, affecting transcription rate, mRNA stability, and mRNA translation into functional proteins. What was once believed to be a specialized function occurring only in plants [8,9] or worms [1,10] has now been shown to control regulatory pathways in several mammalian biological processes, such as cell growth and apoptosis [11][12][13][14], viral infection [15][16][17][18][19], neural function [20 -23], and stem cell function [21, 24 -37]. Here, we outline the issues related to microRNA expression profiling in MSC as an example of how these studies are useful for classifying differentiating cells and, potentially, understanding molecular networks active during differentiation.…”

Section: Introductionmentioning

confidence: 99%

Concise Review: MicroRNA Expression in Multipotent Mesenchymal Stromal Cells

Lakshmipathy¹,

Hart

2007

Stem Cells

118

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Mesenchymal stem cells, or multipotent mesenchymal stromal cells (MSC), isolated from various adult tissue sources have the capacities to self-renew and to differentiate into multiple lineages. Both of these processes are tightly regulated by genetic and epigenetic mechanisms. Emerging evidence indicates that the class of single-stranded noncoding RNAs known as microRNAs also plays a critical role in this process. First described in nematodes and plants, microRNAs have been shown to modulate major regulatory mechanisms in eukaryotic cells involved in a broad array of cellular functions. Studies with various types of embryonic as well as adult stem cells indicate an intricate network of microRNAs regulating key transcription factors and other genes, which in turn determine cell fate. In addition, expression of unique microRNAs in specific cell types serves as a useful diagnostic marker to define a particular cell type. MicroRNAs are also found to be regulated by extracellular signaling pathways that are important for differentiation into specific tissues, suggesting that they play a role in specifying tissue identity. In this review, we describe the importance of microRNAs in stem cells, focusing on our current understanding of microRNAs in MSC and their derivatives.

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“…22 However, we and others recognized that a high level of sustained shRNA expression may be toxic to cells because of competition with endogenous micro-RNA biogenesis, induction of interferon responses, and/or off-targeting effects. 23,[26][27][28][29][30][31][32][33] To stably reduce CCR5 expression without cytotoxicity, we identified a highly efficient shRNA (shRNA 1005) directed to human CCR5 mRNA using the enzymatic production of RNAi libraries (EPRIL) screening technique. 21,34 We expressed shRNA 1005 using the transcriptionally weak H1 promoter to stably reduce CCR5 expression without inducing cytotoxicity in human primary peripheral blood lymphocytes in vitro.…”

Section: Introductionmentioning

confidence: 99%

A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model

Shimizu

Hong

Balamurugan

et al. 2010

Blood

136

166

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IntroductionChemokine receptor CCR5 is an attractive therapeutic target for inhibiting HIV-1, as it serves as a HIV-1 coreceptor and is essential for CCR5 tropic HIV-1 infection. [1][2][3][4] Blocking CCR5 expression should prevent HIV-1 infection at the initial stage of the viral life cycle. Individuals with a ⌬32/⌬32 homozygous mutation in the CCR5 gene do not express CCR5, are highly protected from HIV-1, and are apparently normal. [5][6][7] Recently, an HIV ϩ acute myelogenous leukemia patient was treated for leukemia and HIV infection by bone marrow transplantation using donated CCR5 ⌬32/⌬32 marrow. After the transplantation, nearly 100% of the patient's blood cells were replaced with donor cells. HIV DNA and RNA were undetectable at 20 months, even after the discontinuation of highly active antiretroviral therapy. 8 This evidence supports that long-term and stable reduction of CCR5 is a promising strategy for treating HIV-infected patients. The major limitation of this strategy is the difficulty of identifying human leukocyte antigen-matched CCR5 ⌬32/⌬32 homozygous donors as the mutation exists in approximately 1% of white populations and is rare in other ethnic populations. 9 Small interfering RNAs (siRNAs) induce sequence-specific degradation of mRNAs by RNA interference. 10 Many forms of siRNA have been used to inhibit HIV coreceptors and HIV-1 gene expression in in vitro and in vivo experimental settings. [11][12][13][14][15][16][17][18] To stably inhibit HIV replication, we and others developed lentiviral vectors that are capable of stably delivering short hairpin RNA (shRNA) in mammalian cells. [19][20][21][22][23][24][25] We demonstrated that expression of CCR5-specific shRNA in human primary T lymphocytes results in efficient CCR5-knockdown and protection of cells from HIV-1 infection in vitro. 22 However, we and others recognized that a high level of sustained shRNA expression may be toxic to cells because of competition with endogenous micro-RNA biogenesis, induction of interferon responses, and/or off-targeting effects. 23,[26][27][28][29][30][31][32][33] To stably reduce CCR5 expression without cytotoxicity, we identified a highly efficient shRNA (shRNA 1005) directed to human CCR5 mRNA using the enzymatic production of RNAi libraries (EPRIL) screening technique. 21,34 We expressed shRNA 1005 using the transcriptionally weak H1 promoter to stably reduce CCR5 expression without inducing cytotoxicity in human primary peripheral blood lymphocytes in vitro. 21,34 To test stable CCR5 reduction in vivo, we used a nonhuman primate hematopoietic stem cell transplantation model in which we were able to demonstrate stable reduction of CCR5 expression in peripheral blood lymphocytes in shRNA-transduced CD34 ϩ cell-transplanted rhesus macaques. 21 Because of a single nucleotide mismatch in the shRNA 1005 target sequence between human and rhesus macaque CCR5 mRNA, we mutated the human CCR5 shRNA 1005 so that it would be 100% homologous to the corresponding rhesus macaque CCR5 mRNA target sequence. This rhes...

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Inhibition of HIV-1 replication with designed miRNAs expressed from RNA polymerase II promoters

Cited by 27 publications

References 38 publications

Gamma-Retroviral Vector Design for the Co-Expression of Artificial microRNAs and Therapeutic Proteins

Gamma-Retroviral Vector Design for the Co-Expression of Artificial microRNAs and Therapeutic Proteins

Concise Review: MicroRNA Expression in Multipotent Mesenchymal Stromal Cells

A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model

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