Macromonas sp. C15
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA612959 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:H1-HeLa cells were stably transduced with lentiCas9-Blast (Addgene, Plasmid #52962) and subsequently selected using blasticidin to generate constitutively expressing Cas9 H1-HeLa cells. A single Cas9-expressing H1-HeLa clone was then transduced with lentivirus without a selection marker to stably express CDHR3 C529Y (H1-HeLa+CDHR3). A single CDHR3-expressing H1-HeLa clone was then chosen based on RT-qPCR of CHDR3 expression and RV-C15 RNA levels for mutagenesis. 300 million of the H1-HeLa cells constitutively expressing CDHR3 and Cas9 were transduced with the lentiGuide-Puro from the GeCKO v2 library at a MOI of 0.3. Cells were selected using puromycin and heterogeneous H1-HeLa knockout cell populations were subsequently pooled together. The CRISPR genetic screens were started 10 days post transduction. >1000-fold coverage of mutagenized cells (libraries A and B) was infected with either RV-C15 (MOI=1 PFU/cell) or EV-D68 Missouri (MOI=1 PFU/cell). RV-C15 infection was repeated for an additional round at 6 days post-infection. As soon as appearance of visibly viable colonies was observed, populations of virus-resistant cells were pooled and harvested. Uninfected starting populations of mutagenized cells were used as the unselected reference. Total genomic DNA from both virus-resistant and uninfected cells was respectively extracted using QIAamp DNA Mini Kit (Qiagen). The inserted guide RNA sequences were retrieved from the genomic DNA by PCR amplification. The PCR products were then purified and subjected to NextSeq platform (Illumina) next-generation sequencing.
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Project description:Pseudomonas syringae strain:C15 Genome sequencing and assembly
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Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD8+ effector (CD62L-CD44+) T cell development, we performed bulk RNA-seq on splenic CD8+ effector T cells 7 days post-infection with either WT ECTV or ECTV∆C15.
2026-03-12 | GSE308808 | GEO
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD4+ effector (CD62L-CD44+) T cell development, we performed bulk RNA-seq on splenic CD4+ effector T cells 7 days post-infection with either WT ECTV or ECTV∆C15.
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Project description:Euryarchaeota archaeon JGI 0000059-C15 Genome sequencing
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Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. Using the model poxvirus, ectromelia (ECTV), our lab has demonstrated that its B22 protein, C15, is capable of inhibiting both CD4+ and CD8+ T cells both in vitro and ex vivo, similar to other B22 proteins. Furthermore, we have also identified an additional function of C15: antagonism of NK cells during early infection. Specifically, C15 antagonizes the engagement of NK cells with infected cells in the popliteal lymph node during the innate immune response to infection. This inhibition by C15 promotes viral dissemination and replication, such that in the absence of C15, viral replication is significantly stunted. To date, there has not been any work investigating the mechanisms underlying this NK cell inhibition. To broadly explore the transcriptional changes occurring during this critical window, we performed bulk RNA-seq on popliteal lymph nodes at 48 hours post-infection.
2026-03-12 | GSE309219 | GEO
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. Using the model poxvirus, ectromelia (ECTV), our lab has demonstrated that its B22 protein, C15, is capable of inhibiting both CD4+ and CD8+ T cells both in vitro and ex vivo, similar to other B22 proteins. Furthermore, we have also identified an additional function of C15: antagonism of NK cells during early infection. Specifically, C15 antagonizes the engagement of NK cells with infected cells in the popliteal lymph node during the innate immune response to infection. This inhibition by C15 promotes viral dissemination and replication, such that in the absence of C15, viral replication is significantly stunted. To date, there has not been any work investigating the mechanisms underlying this NK cell inhibition. To broadly explore the cell type-specific changes occurring during this critical window, we performed single cell RNA-seq on popliteal lymph nodes at 24 and 48 hours post-infection.
2026-03-12 | GSE309220 | GEO
Project description:Orthopoxviruses, such as variola and Mpox, encode a myriad of immunomodulatory proteins to promote pathogenesis. One notable family is the B22 family of proteins, which are highly conserved surface glycoproteins that potently inhibit T cell activation in vitro and ex vivo. However, there has been limited work investigating the impact of B22 proteins on both CD4+ and CD8+ T cell responses in vivo. Therefore, we used ectromelia (ECTV) as a natural host-pathogen model to investigate the impact of its B22 protein, C15, on T cell responses in vivo. To broadly investigate the impact of C15 on CD8+ effector (CD62L-CD44+) T cell development during the contraction phase, we performed bulk RNA-seq on splenic CD8+ effector T cells 10 days post-infection with either WT ECTV or ECTV∆C15.
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