Efficient inhibition of human cytomegalovirus UL122 gene expression in cell by small interfering RNAs

Duan, Qun-Jun; Tao, Ran; Hu, Miaofeng; Shang, Shuai

doi:10.1002/jobm.200800364

Cited by 5 publications

(3 citation statements)

References 31 publications

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“…To our knowledge, no study has considered using one siRNA to simultaneously abolish the expression of multiple IE genes, even though reports have demonstrated that siRNA-mediated targeting of other open reading frames (ORFs) can affect HCMV replication (4,24,68,74,79). A result of this present study is the demonstration that targeting a conserved and shared region of IE genes can inhibit HCMV replication, thereby illustrating the potential of using this region as a new drug target for RNAi-based therapeutics.…”

Section: Discussionmentioning

confidence: 99%

“…With the ongoing efforts in RNAi-based therapeutic development, small interfering RNAs (siRNAs) and their derivatives have been designed to inhibit the expression of several genes related to virus infections in humans, including those with human immunodeficiency virus type 1, hepatitis C virus, hepatitis B virus, poliovirus, human papillomavirus, and influenza virus (9,31,32,39,52,81), which provides evidence that RNAi has the potential to be an effective strategy to control viral diseases. There are also reports on RNAi-based targeting of HCMV genes (4,22,24,68,74,79).…”

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confidence: 99%

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RNA Interference-Mediated Targeting of Human Cytomegalovirus Immediate-Early or Early Gene Products Inhibits Viral Replication with Differential Effects on Cellular Functions

Xiaofei

Stadler

Debatis

et al. 2012

J Virol

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Viral drug toxicity, resistance, and an increasing immunosuppressed population warrant continued research into new avenues for limiting diseases associated with human cytomegalovirus (HCMV). In this study, a small interfering RNA (siRNA), siX3, was designed to target coding sequences within shared exon 3 of UL123 and UL122 transcripts encoding IE1 and IE2 immediate-early proteins of HCMV. Pretreatment of cells with siX3 reduced the levels of viral protein expression, DNA replication, and progeny virus production compared to control siRNA. Two siRNAs against UL54 and overlapping transcripts ( UL55-57 ) were compared to siX3 in HCMV infection and were also found to be effective at inhibiting HCMV replication. Further investigation into the effects of the siRNAs on viral replication showed that pretreatment with each of the siRNAs resulted in an inhibition in the formation of mature replication compartments. The ability of these siRNAs to prevent or reduce certain cytopathic effects associated with HCMV infection was also examined. Infected cells pretreated with siX3, but not siUL54, retained promyelocytic leukemia (PML) protein in cellular PML bodies, an essential component of this host intrinsic antiviral defense. DNA damage response proteins, which are localized in nuclear viral replication compartments, were reduced in the siX3- and siUL54-treated cells. siX3, but not siUL54, prevented DNA damage response signaling early after infection. Therapeutic efficacy was demonstrated by treating cells with siRNAs after HCMV replication had commenced. Together, these findings suggest that siRNAs targeting exon 3 of the major IE genes or the UL54-57 transcripts be further studied for their potential development into anti-HCMV therapeutics.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

RNA Interference-Mediated Targeting of Human Cytomegalovirus Immediate-Early or Early Gene Products Inhibits Viral Replication with Differential Effects on Cellular Functions

Xiaofei

Stadler

Debatis

et al. 2012

J Virol

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“…The ΔCq values (Cq gene -Cq β-Actin ) were calculated, following which the ΔΔCq (ΔCq treated -ΔCq control ) were calculated. The relative numbers of HCMV DNAs were described using the equation 2 -ΔΔCq (22). The experiments were repeated three times.…”

Section: Stable Expression Of Nudt14 and Transfection Of Nudt14-specimentioning

confidence: 99%

Human cytomegalovirus RL13 protein interacts with host NUDT14 protein affecting viral DNA replication

Wang

Ren

Cui

et al. 2016

Molecular Medicine Reports

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The interaction between the host and human cytomegalovirus (HCMV) is important in determining the outcome of a viral infection. The HCMV RL13 gene product exerts independent, inhibitory effects on viral growth in fibroblasts and epithelial cells. At present, there are few reports on the interactions between the HCMV RL13 protein and human host proteins. The present study provided direct evidence for the specific interaction between HCMV RL13 and host nucleoside diphosphate linked moiety X (nudix)‑type motif 14 (NUDT14), a UDP‑glucose pyrophosphatase, using two‑hybrid screening, an in vitro glutathione S‑transferase pull‑down assay, and co‑immunoprecipitation in human embryonic kidney HEK293 cells. Additionally, the RL13 protein was shown to co‑localize with the NUDT14 protein in the HEK293 cell membrane and cytoplasm, demonstrated using fluorescence confocal microscopy. Decreasing the expression level of NUDT14 via NUDT14‑specific small interfering RNAs increased the number of viral DNA copies in the HCMV‑infected cells. However, the overexpression of NUDT14 in a stably expressing cell line did not affect viral DNA levels significantly in the HCMV infected cells. Based on the known functions of NUDT14, the results of the present study suggested that the interaction between the RL13 protein and NUDT14 protein may be involved in HCMV DNA replication, and that NUDT14 may offer potential in the modulation of viral infection.

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Comparison of inhibitory efficacy of short interfering RNAs targeting different genes on Measles virus replication

Shi

Wang

et al. 2011

J. Basic Microbiol.

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RNA interference (RNAi)-based short interfering RNAs (siRNAs) targeting the viral genes and the host cellular genes have been used to suppress Measles virus (MV) replication in vitro. In order to select suitable target genes and highly effective target sites for developing effective RNAi-mediated anti-MV therapy, in this study, nine short hairpin RNA (shRNA) expression vectors, which expressed siRNAs targeting the host celluar Rab9 GTPase gene, the viral large protein (L) gene and nucleoprotein (N) gene, respectively, were constructed and used to compare their ability to inhibit MV replication in Vero-E6 cells. The results showed that nine siRNAs targeting different genes exhibited different inhibitory efficacy on MV replication in vitro (about 23-94%), which could last at least 168 h post-infection. Of the nine siRNAs, R2, L1 and N2 more effectively decreased MV replication by over 90%. Furthermore, inhibitory efficacy on MV replication were increased and reached almost 100% when cells were transfected with pR2, pL1 and pN2 together. These results emphasis the importance of selecting suitable siRNA target sites for developing siRNAs-based drug therapy for MV, and demonstrate the potential of combination of siRNAs targeting different genes as a therapeutic approach to treat MV infection.

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Efficient inhibition of human cytomegalovirus UL122 gene expression in cell by small interfering RNAs

Cited by 5 publications

References 31 publications

RNA Interference-Mediated Targeting of Human Cytomegalovirus Immediate-Early or Early Gene Products Inhibits Viral Replication with Differential Effects on Cellular Functions

RNA Interference-Mediated Targeting of Human Cytomegalovirus Immediate-Early or Early Gene Products Inhibits Viral Replication with Differential Effects on Cellular Functions

Human cytomegalovirus RL13 protein interacts with host NUDT14 protein affecting viral DNA replication

Comparison of inhibitory efficacy of short interfering RNAs targeting different genes on Measles virus replication

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