Abhishek N Prasad
I am currently (as of October 2014) a postdoctoral fellow at Galveston National Laboratory, part of the University of Texas Medical Branch (UTMB) in Galveston, TX. My research interests include emerging viral pathogens, innate immune responses to virus infection, host-pathogen interaction, and viral immune evasion.
Currently, I am developing projects investigating the comparitive immunology of humans and fruit bats (specifically Rousettus aegyptiacus) to filovirus (Marburg, Ebola) infection.
As a graduate student beginning at the University of New Mexico School of Medicine (Dept. of Pathology), and continuing on to Colorado State University College of Veterinary Medicine and Biomedical Sciences (Dept. of Microbiology, Immunology, and Pathology), my Ph.D research involved studying mosquito small RNA pathways in the context of innate immunity to arthropod-borne viruses (arboviruses). Most of my work revolved around using a Culex spp. mosquito/West Nile virus (Flaviviridae) model. My approach to this research involved the use of next-generation sequencing technology to profile the anti-WNV small-interfering RNA (siRNA) and PIWI-interacting RNA (piRNA) pathway responses after infection with WNV and other arboviruses (Western equine encephalitis virus (WEEV) and Sindbis virus (SINV), both alphaviruses). I successfully defended my PhD dissertation in June 2014.
Supervisors: Gregory Ebel and Alexander Bukreyev
Currently, I am developing projects investigating the comparitive immunology of humans and fruit bats (specifically Rousettus aegyptiacus) to filovirus (Marburg, Ebola) infection.
As a graduate student beginning at the University of New Mexico School of Medicine (Dept. of Pathology), and continuing on to Colorado State University College of Veterinary Medicine and Biomedical Sciences (Dept. of Microbiology, Immunology, and Pathology), my Ph.D research involved studying mosquito small RNA pathways in the context of innate immunity to arthropod-borne viruses (arboviruses). Most of my work revolved around using a Culex spp. mosquito/West Nile virus (Flaviviridae) model. My approach to this research involved the use of next-generation sequencing technology to profile the anti-WNV small-interfering RNA (siRNA) and PIWI-interacting RNA (piRNA) pathway responses after infection with WNV and other arboviruses (Western equine encephalitis virus (WEEV) and Sindbis virus (SINV), both alphaviruses). I successfully defended my PhD dissertation in June 2014.
Supervisors: Gregory Ebel and Alexander Bukreyev
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Small RNA (sRNA) regulatory pathways (SRRPs) are important to anti-viral defence in mosquitoes. To identify critical features of the virus infection process in Dengue serotype 2 (DENV2)-infected Ae. aegypti, we deep-sequenced small non-coding RNAs. Triplicate biological replicates were used so that rigorous statistical metrics could be applied.
Results
In addition to virus-derived siRNAs (20-23 nts) previously reported for other arbovirus-infected mosquitoes, we show that PIWI pathway sRNAs (piRNAs) (24-30 nts) and unusually small RNAs (usRNAs) (13-19 nts) are produced in DENV-infected mosquitoes. We demonstrate that a major catalytic enzyme of the siRNA pathway, Argonaute 2 (Ago2), co-migrates with a ~1 megadalton complex in adults prior to bloodfeeding. sRNAs were cloned and sequenced from Ago2 immunoprecipitations. Viral sRNA patterns change over the course of infection. Host sRNAs were mapped to the published aedine transcriptome and subjected to analysis using edgeR (Bioconductor). We found that sRNA profiles are altered early in DENV2 infection, and mRNA targets from mitochondrial, transcription/translation, and transport functional categories are affected. Moreover, small non-coding RNAs (ncRNAs), such as tRNAs, spliceosomal U RNAs, and snoRNAs are highly enriched in DENV-infected samples at 2 and 4 dpi.
Conclusions
These data implicate the PIWI pathway in anti-viral defense. Changes to host sRNA profiles indicate that specific cellular processes are affected during DENV infection, such as mitochondrial function and ncRNA levels. Together, these data provide important progress in understanding the DENV2 infection process in Ae. aegypti."
Small RNA (sRNA) regulatory pathways (SRRPs) are important to anti-viral defence in mosquitoes. To identify critical features of the virus infection process in Dengue serotype 2 (DENV2)-infected Ae. aegypti, we deep-sequenced small non-coding RNAs. Triplicate biological replicates were used so that rigorous statistical metrics could be applied.
Results
In addition to virus-derived siRNAs (20-23 nts) previously reported for other arbovirus-infected mosquitoes, we show that PIWI pathway sRNAs (piRNAs) (24-30 nts) and unusually small RNAs (usRNAs) (13-19 nts) are produced in DENV-infected mosquitoes. We demonstrate that a major catalytic enzyme of the siRNA pathway, Argonaute 2 (Ago2), co-migrates with a ~1 megadalton complex in adults prior to bloodfeeding. sRNAs were cloned and sequenced from Ago2 immunoprecipitations. Viral sRNA patterns change over the course of infection. Host sRNAs were mapped to the published aedine transcriptome and subjected to analysis using edgeR (Bioconductor). We found that sRNA profiles are altered early in DENV2 infection, and mRNA targets from mitochondrial, transcription/translation, and transport functional categories are affected. Moreover, small non-coding RNAs (ncRNAs), such as tRNAs, spliceosomal U RNAs, and snoRNAs are highly enriched in DENV-infected samples at 2 and 4 dpi.
Conclusions
These data implicate the PIWI pathway in anti-viral defense. Changes to host sRNA profiles indicate that specific cellular processes are affected during DENV infection, such as mitochondrial function and ncRNA levels. Together, these data provide important progress in understanding the DENV2 infection process in Ae. aegypti."