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Article: Maxizyme-mediated suppression of chikungunya virus replication and transmission in transgenic

Mishra, Priya / Balaraman, Velmurugan / Fraser, Malcolm J

2023 Volume 14, Page(s) 1286519

Abstract: Chikungunya virus (CHIKV) is an emerging mosquito-borne pathogen of significant public health importance. There are currently no prophylactic vaccines or therapeutics available to control CHIKV. One approach to arbovirus control that has been proposed is ...

Abstract	Chikungunya virus (CHIKV) is an emerging mosquito-borne pathogen of significant public health importance. There are currently no prophylactic vaccines or therapeutics available to control CHIKV. One approach to arbovirus control that has been proposed is the replacement of transmission-competent mosquitoes with those that are refractory to virus infection. Several transgene effectors are being examined as potentially useful for this population replacement approach. We previously demonstrated the successful use of hammerhead ribozymes (hRzs) as an antiviral effector transgene to control CHIKV infection of, and transmission by, Aedes mosquitoes. In this report we examine a maxizyme approach to enhance the catalytic activity and prevent virus mutants from escaping these ribozymes. We designed a maxizyme containing minimized (monomer) versions of two hRzs we previously demonstrated to be the most effective in CHIKV suppression. Three versions of CHIKV maxizyme were designed: Active (Mz), inactive (ΔMz), and a connected CHIKV maxizyme (cMz). The maxizymes with their expression units (Ae-tRNA
Language	English
Publishing date	2023-12-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2023.1286519
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Article ; Online: RT-qPCR genotyping assays for differentiating Rift Valley fever phlebovirus strains

Balaraman, Velmurugan / Gaudreault, Natasha N. / Trujillo, Jessie D. / Indran, Sabarish V. / Wilson, William C. / Richt, Juergen A.

Elsevier B.V. Journal of Virological Methods. 2023 May, v. 315 p.114693-

2023

Abstract: Rift Valley fever phlebovirus (RVFV) is an emerging, mosquito-borne, zoonotic pathogen. Real time RT-qPCR genotyping (GT) assays were developed to differentiate between two RVFV wild-type strains (128B-15 and SA01–1322) and a vaccine strain (MP-12). The ... ...

Abstract	Rift Valley fever phlebovirus (RVFV) is an emerging, mosquito-borne, zoonotic pathogen. Real time RT-qPCR genotyping (GT) assays were developed to differentiate between two RVFV wild-type strains (128B-15 and SA01–1322) and a vaccine strain (MP-12). The GT assay uses a one-step RT-qPCR mix, with two different RVFV strain-specific primers (either forward or reverse) with long or short G/C tags and a common primer (either forward or reverse) for each of the 3 genomic segments. The GT assay produces PCR amplicons with unique melting temperatures that are resolved in a post PCR melt curve analysis for strain identification. Furthermore, a strain specific RT-qPCR (SS-PCR) assay was developed to allow for specific detection of low titer RVFV strains in mixed RVFV samples. Our data shows that the GT assays are capable of differentiating L, M, and S segments of RVFV strains 128B-15 versus MP-12, and 128B-15 versus SA01–1322. The SS-PCR assay results revealed that it can specifically amplify and detect a low titer MP-12 strain in mixed RVFV samples. Overall, these two novel assays are useful as screening tools for determining reassortment of the segmented RVFV genome during co-infections, and could be adapted and applied for other segmented pathogens of interest.
Keywords	Rift Valley fever phlebovirus ; genome ; genomics ; genotyping ; pathogens ; vaccines ; Reassortment ; Genotyping assays ; Strain-specific RT-PCR ; Melt curve analysis
Language	English
Dates of publication	2023-05
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2023.114693
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Article ; Online: RT-qPCR genotyping assays for differentiating Rift Valley fever phlebovirus strains.

Balaraman, Velmurugan / Gaudreault, Natasha N / Trujillo, Jessie D / Indran, Sabarish V / Wilson, William C / Richt, Juergen A

Journal of virological methods

2023 Volume 315, Page(s) 114693

Abstract	Rift Valley fever phlebovirus (RVFV) is an emerging, mosquito-borne, zoonotic pathogen. Real time RT-qPCR genotyping (GT) assays were developed to differentiate between two RVFV wild-type strains (128B-15 and SA01-1322) and a vaccine strain (MP-12). The GT assay uses a one-step RT-qPCR mix, with two different RVFV strain-specific primers (either forward or reverse) with long or short G/C tags and a common primer (either forward or reverse) for each of the 3 genomic segments. The GT assay produces PCR amplicons with unique melting temperatures that are resolved in a post PCR melt curve analysis for strain identification. Furthermore, a strain specific RT-qPCR (SS-PCR) assay was developed to allow for specific detection of low titer RVFV strains in mixed RVFV samples. Our data shows that the GT assays are capable of differentiating L, M, and S segments of RVFV strains 128B-15 versus MP-12, and 128B-15 versus SA01-1322. The SS-PCR assay results revealed that it can specifically amplify and detect a low titer MP-12 strain in mixed RVFV samples. Overall, these two novel assays are useful as screening tools for determining reassortment of the segmented RVFV genome during co-infections, and could be adapted and applied for other segmented pathogens of interest.
MeSH term(s)	Animals ; Humans ; Rift Valley Fever/diagnosis ; Rift Valley fever virus/genetics ; Genotype ; Phlebovirus ; Polymerase Chain Reaction
Language	English
Publishing date	2023-02-16
Publishing country	Netherlands
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2023.114693
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Article: Preparation of an Intelligent pH Film Based on Biodegradable Polymers for Monitoring the Food Quality and Reducing the Microbial Contaminants.

Subramanian, Kumaran / Balaraman, Deivasigamani / Kaliyaperumal, Kumaravel / Devi Rajeswari, V / Balakrishnan, K / Ronald Ross, P / Perumal, Elumalai / Sampath Renuga, Pugazhvendan / Panangal, Mani / Swarnalatha, Y / Velmurugan, S

publication RETRACTED

Bioinorganic chemistry and applications

2022 Volume 2022, Page(s) 7975873

Abstract: Hydrogel refers to a three-dimensional cross-linked polymeric network made of synthetic or natural polymers that can hold water in its porous structure. The inclusion of hydrophilic groups in the polymer chains, such as amino, carboxyl, and hydroxyl ... ...

Abstract	Hydrogel refers to a three-dimensional cross-linked polymeric network made of synthetic or natural polymers that can hold water in its porous structure. The inclusion of hydrophilic groups in the polymer chains, such as amino, carboxyl, and hydroxyl groups, contributes to the hydrogel's water-holding ability. At physiological temperature and pH, these polymeric materials do not dissolve in water, but they do swell significantly in aqueous media. Hydrogel can be manufactured out of almost any water-soluble polymer, and it comes in a variety of chemical compositions and bulk physical properties. Hydrogel can also be made in a variety of ways. Hydrogel comes in a variety of physical shapes, including slabs, microparticles, nanoparticles, coatings, and films. Due to its ease of manufacture and self-application in clinical and fundamental applications, hydrogel has been widely exploited as a drug carrier. Contact lenses, artificial corneas, wound dressing, suture coating, catheters, and electrode sensors are some of the biomedical applications of hydrogels. The pigment color changes were observed from colorless to pale pink followed by dark reddish-pink. Anthocyanin was produced in large quantities and tested using a UV-visible spectrophotometer. At 450-550 nm, the largest peak (absorbance) was detected, indicating the presence of anthocyanin. The FTIR analysis of this study shows the different stretches of bonds at different peaks: 2918.309 (-C-H alkane stretch), 2812.12 (-C-H aldehyde weak intensity), 192320.37/cm (C-O bend), 21915.50, 2029.08/cm (-C=C arene group), 1906.94/cm (=C-H aromatics), 1797.78/cm (=C-H), 1707.94 (-C=O ketene), 1579.70, 1382.96 (C-H alkane strong bend), 889.18/cm (C-H aromatics plane bend), and 412.77/cm (-C-CI strong bond). The spectra of the PVA/chitosan film depict the peak's formation: 1571.88, 1529.55, 1500.62/cm (C-H alkene strong bend), 1492.90, 1483.26, 1467.83/cm (C-H alkene strong bond), 670.48, 443.63, 412.77/cm (-O-H carboxylic acids with great intensity), 1708.93 (-C=O ketone), and 1656.0/cm (alkenyl C=C stretch strong bond).
Language	English
Publishing date	2022-06-20
Publishing country	Egypt
Document type	Journal Article ; Retracted Publication
ZDB-ID	2213020-2
ISSN	1687-479X ; 1565-3633
ISSN (online)	1687-479X
ISSN	1565-3633
DOI	10.1155/2022/7975873
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Article: Identification of host factors for Rift Valley Fever Phlebovirus.

Balaraman, Velmurugan / Indran, Sabarish V / Li, Yonghai / Meekins, David A / Jakkula, Laxmi U M R / Liu, Heidi / Hays, Micheal P / Souza-Neto, Jayme A / Gaudreault, Natasha N / Hardwidge, Philip R / Wilson, William C / Weber, Friedemann / Richt, Juergen A

bioRxiv : the preprint server for biology

2023

Abstract: Background: Rift Valley fever phlebovirus (RVFV) is a zoonotic pathogen that causes Rift Valley fever (RVF) in livestock and humans. Currently, there is no licensed human vaccine or antiviral drug to control RVF. Although multiple species of animals and ...

Abstract	Background: Rift Valley fever phlebovirus (RVFV) is a zoonotic pathogen that causes Rift Valley fever (RVF) in livestock and humans. Currently, there is no licensed human vaccine or antiviral drug to control RVF. Although multiple species of animals and humans are vulnerable to RVFV infection, host factors affecting susceptibility are not well understood. Methodology: To identify the host factors or genes essential for RVFV replication, we conducted a CRISPR-Cas9 knock-out screen in human A549 cells. We then validated the putative genes using siRNA-mediated knockdowns and CRISPR-Cas9-mediated knockout studies, respectively. The role of a candidate gene in the virus replication cycle was assessed by measuring intracellular viral RNA accumulation, and the virus titers by plaque assay or TCID Findings: We identified approximately 900 genes with potential involvement in RVFV infection and replication. Further evaluation of the effect of six genes on viral replication using siRNA-mediated knockdowns found that silencing two genes (WDR7 and LRP1) significantly impaired RVFV replication. For further analysis, we focused on the Conclusion: In summary, we have identified
Language	English
Publishing date	2023-09-29
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.09.28.559935
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Article ; Online: Identification of Host Factors for Rift Valley Fever Phlebovirus.

Viruses

2023 Volume 15, Issue 11

Abstract: Rift Valley fever phlebovirus (RVFV) is a zoonotic pathogen that causes Rift Valley fever (RVF) in livestock and humans. Currently, there is no licensed human vaccine or antiviral drug to control RVF. Although multiple species of animals and humans are ... ...

Abstract	Rift Valley fever phlebovirus (RVFV) is a zoonotic pathogen that causes Rift Valley fever (RVF) in livestock and humans. Currently, there is no licensed human vaccine or antiviral drug to control RVF. Although multiple species of animals and humans are vulnerable to RVFV infection, host factors affecting susceptibility are not well understood. To identify the host factors or genes essential for RVFV replication, we conducted CRISPR-Cas9 knockout screening in human A549 cells. We then validated the putative genes using siRNA-mediated knock-downs and CRISPR-Cas9-mediated knock-out studies. The role of a candidate gene in the virus replication cycle was assessed by measuring intracellular viral RNA accumulation, and the virus titers were analyzed using plaque assay or TCID
MeSH term(s)	Animals ; Humans ; Rift Valley Fever ; Rift Valley fever virus/genetics ; Phlebovirus/genetics ; Virus Replication ; RNA, Small Interfering/genetics ; RNA, Small Interfering/pharmacology ; Adaptor Proteins, Signal Transducing
Chemical Substances	RNA, Small Interfering ; WDR7 protein, human ; Adaptor Proteins, Signal Transducing
Language	English
Publishing date	2023-11-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15112251
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Article ; Online: Evaluating α-galactosylceramide as an adjuvant for live attenuated influenza vaccines in pigs.

Artiaga, Bianca L / Morozov, Igor / Ransburgh, Russell / Kwon, Taeyong / Balaraman, Velmurugan / Indran, Sabarish V / De Carvalho Madrid, Darling Melany / Gu, Weihong / Henningson, Jamie / Ma, Wenjun / Richt, Jürgen A / Driver, John P

Animal diseases

2022 Volume 2, Issue 1, Page(s) 19

Abstract: Natural killer T (NKT) cells activated with the glycolipid ligand α-galactosylceramide (α-GalCer) stimulate a wide variety of immune cells that enhance vaccine-mediated immune responses. Several studies have used this approach to adjuvant inactivated and ...

Abstract	Natural killer T (NKT) cells activated with the glycolipid ligand α-galactosylceramide (α-GalCer) stimulate a wide variety of immune cells that enhance vaccine-mediated immune responses. Several studies have used this approach to adjuvant inactivated and subunit influenza A virus (IAV) vaccines, including to enhance cross-protective influenza immunity. However, less is known about whether α-GalCer can enhance live attenuated influenza virus (LAIV) vaccines, which usually induce superior heterologous and heterosubtypic immunity compared to non-replicating influenza vaccines. The current study used the swine influenza challenge model to assess whether α-GalCer can enhance cross-protective immune responses elicited by a recombinant H3N2 LAIV vaccine (TX98ΔNS1) encoding a truncated NS1 protein. In one study, weaning pigs were administered the H3N2 TX98ΔNS1 LAIV vaccine with 0, 10, 50, and 100 μg/kg doses of α-GalCer, and subsequently challenged with a heterologous H3N2 virus. All treatment groups were protected from infection. However, the addition of α-GalCer appeared to suppress nasal shedding of the LAIV vaccine. In another experiment, pigs vaccinated with the H3N2 LAIV, with or without 50 μg/kg of α-GalCer, were challenged with the heterosubtypic pandemic H1N1 virus. Pigs vaccinated with the LAIV alone generated cross-reactive humoral and cellular responses which blocked virus replication in the airways, and significantly decreased virus shedding. On the other hand, combining the vaccine with α-GalCer reduced cross-protective cellular and antibody responses, and resulted in higher virus titers in respiratory tissues. These findings suggest that: (i) high doses of α-GalCer impair the replication and nasal shedding of the LAIV vaccine; and (ii) α-GalCer might interfere with heterosubtypic cross-protective immune responses. This research raise concerns that should be considered before trying to use NKT cell agonists as a possible adjuvant approach for LAIV vaccines. Supplementary information: The online version contains supplementary material available at 10.1186/s44149-022-00051-x.
Language	English
Publishing date	2022-08-01
Publishing country	England
Document type	Journal Article
ISSN	2731-0442
ISSN (online)	2731-0442
DOI	10.1186/s44149-022-00051-x
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Article ; Online: Rift Valley fever virus Gn V5-epitope tagged virus enables identification of UBR4 as a Gn interacting protein that facilitates Rift Valley fever virus production.

Bracci, Nicole / de la Fuente, Cynthia / Saleem, Sahar / Pinkham, Chelsea / Narayanan, Aarthi / García-Sastre, Adolfo / Balaraman, Velmurugan / Richt, Juergen A / Wilson, William / Kehn-Hall, Kylene

Virology

2022 Volume 567, Page(s) 65–76

Abstract: Rift Valley fever virus (RVFV) is an arbovirus that was first reported in the Rift Valley of Kenya which causes significant disease in humans and livestock. RVFV is a tri-segmented, negative-sense RNA virus consisting of a L, M, and S segments with the M ...

Abstract	Rift Valley fever virus (RVFV) is an arbovirus that was first reported in the Rift Valley of Kenya which causes significant disease in humans and livestock. RVFV is a tri-segmented, negative-sense RNA virus consisting of a L, M, and S segments with the M segment encoding the glycoproteins Gn and Gc. Host factors that interact with Gn are largely unknown. To this end, two viruses containing an epitope tag (V5) on the Gn protein in position 105 or 229 (V5Gn105 and V5Gn229) were generated using the RVFV MP-12 vaccine strain as a backbone. The V5-tag insertion minimally impacted Gn functionality as measured by replication kinetics, Gn localization, and antibody neutralization assays. A proteomics-based approach was used to identify novel Gn-binding host proteins, including the E3 ubiquitin-protein ligase, UBR4. Depletion of UBR4 resulted in a significant decrease in RVFV titers and a reduction in viral RNA production.
MeSH term(s)	Animals ; Antibodies, Neutralizing/metabolism ; Antibodies, Viral/metabolism ; Calmodulin-Binding Proteins/genetics ; Calmodulin-Binding Proteins/metabolism ; Cell Line ; Cell Line, Tumor ; Culex ; Epitopes/chemistry ; Epitopes/metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; HEK293 Cells ; Hepatocytes/virology ; Host-Pathogen Interactions/genetics ; Humans ; Protein Binding ; Rift Valley fever virus/genetics ; Rift Valley fever virus/metabolism ; Signal Transduction ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism ; Viral Envelope Proteins/genetics ; Viral Envelope Proteins/metabolism ; Virus Replication
Chemical Substances	Antibodies, Neutralizing ; Antibodies, Viral ; Calmodulin-Binding Proteins ; Epitopes ; Viral Envelope Proteins ; UBR4 protein, human (EC 2.3.2.27) ; Ubiquitin-Protein Ligases (EC 2.3.2.27)
Language	English
Publishing date	2022-01-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	200425-2
ISSN	1096-0341 ; 0042-6822
ISSN (online)	1096-0341
ISSN	0042-6822
DOI	10.1016/j.virol.2021.12.010
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Article ; Online: Preparation of an Intelligent pH Film Based on Biodegradable Polymers for Monitoring the Food Quality and Reducing the Microbial Contaminants

Kumaran Subramanian / Deivasigamani Balaraman / Kumaravel Kaliyaperumal / V. Devi Rajeswari / K. Balakrishnan / P. Ronald Ross / Elumalai Perumal / Pugazhvendan Sampath Renuga / Mani Panangal / Y. Swarnalatha / S. Velmurugan

Bioinorganic Chemistry and Applications, Vol

2022 Volume 2022

Abstract	Hydrogel refers to a three-dimensional cross-linked polymeric network made of synthetic or natural polymers that can hold water in its porous structure. The inclusion of hydrophilic groups in the polymer chains, such as amino, carboxyl, and hydroxyl groups, contributes to the hydrogel’s water-holding ability. At physiological temperature and pH, these polymeric materials do not dissolve in water, but they do swell significantly in aqueous media. Hydrogel can be manufactured out of almost any water-soluble polymer, and it comes in a variety of chemical compositions and bulk physical properties. Hydrogel can also be made in a variety of ways. Hydrogel comes in a variety of physical shapes, including slabs, microparticles, nanoparticles, coatings, and films. Due to its ease of manufacture and self-application in clinical and fundamental applications, hydrogel has been widely exploited as a drug carrier. Contact lenses, artificial corneas, wound dressing, suture coating, catheters, and electrode sensors are some of the biomedical applications of hydrogels. The pigment color changes were observed from colorless to pale pink followed by dark reddish-pink. Anthocyanin was produced in large quantities and tested using a UV-visible spectrophotometer. At 450–550 nm, the largest peak (absorbance) was detected, indicating the presence of anthocyanin. The FTIR analysis of this study shows the different stretches of bonds at different peaks: 2918.309 (-C-H alkane stretch), 2812.12 (-C-H aldehyde weak intensity), 192320.37/cm (C-O bend), 21915.50, 2029.08/cm (-C=C arene group), 1906.94/cm (=C-H aromatics), 1797.78/cm (=C-H), 1707.94 (-C=O ketene), 1579.70, 1382.96 (C-H alkane strong bend), 889.18/cm (C-H aromatics plane bend), and 412.77/cm (-C-CI strong bond). The spectra of the PVA/chitosan film depict the peak’s formation: 1571.88, 1529.55, 1500.62/cm (C-H alkene strong bend), 1492.90, 1483.26, 1467.83/cm (C-H alkene strong bond), 670.48, 443.63, 412.77/cm (-O-H carboxylic acids with great intensity), 1708.93 (-C=O ketone), ...
Keywords	Biotechnology ; TP248.13-248.65 ; Inorganic chemistry ; QD146-197
Subject code	540
Language	English
Publishing date	2022-01-01T00:00:00Z
Publisher	Hindawi Limited
Document type	Article ; Online
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Article: Disruption of dengue virus transmission by mosquitoes.

Franz, Alexander W E / Balaraman, Velmurugan / Fraser, Malcolm J

Current opinion in insect science

2015 Volume 8, Page(s) 88–96

Abstract: Current control efforts for mosquito-borne arboviruses focus on mosquito control involving insecticide applications, which are becoming increasingly ineffective and unsustainable in urban areas. Mosquito population replacement is an alternative arbovirus ...

Abstract	Current control efforts for mosquito-borne arboviruses focus on mosquito control involving insecticide applications, which are becoming increasingly ineffective and unsustainable in urban areas. Mosquito population replacement is an alternative arbovirus control concept aiming at replacing virus-competent vector populations with laboratory-engineered incompetent vectors. A prerequisite for this strategy is the design of robust anti-pathogen effectors that can ultimately be genetically driven through a wild-type population. Several anti-pathogen effector concepts have been developed that target the RNA genomes of arboviruses such as dengue virus in a highly sequence-specific manner. Design principles are based on long inverted-repeat RNA triggered RNA interference, catalytic hammerhead ribozymes, and trans-splicing Group I Introns that are able to induce apoptosis in virus-infected cells following splicing with target viral RNA.
Language	English
Publishing date	2015-04-01
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2772833-X
ISSN	2214-5753 ; 2214-5745
ISSN (online)	2214-5753
ISSN	2214-5745
DOI	10.1016/j.cois.2014.12.009
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