Genome Wide:Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-Rad S3e sorting.
Article Title: The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8 + T Cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796 ; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-rad S3e sorting.
Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells.
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2one vector system (co-expressing spCas9and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench54) and prepared in the Yale Cancer Center Functional Genomics core). pMSCVU6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatiblewithBio-RadS3e sorting.
CRISPR:Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-Rad S3e sorting.
Article Title: The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8 + T Cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796 ; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-rad S3e sorting.
Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells.
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2one vector system (co-expressing spCas9and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench54) and prepared in the Yale Cancer Center Functional Genomics core). pMSCVU6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatiblewithBio-RadS3e sorting.
Knock-Out:Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-Rad S3e sorting.
Article Title: The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8 + T Cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796 ; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-rad S3e sorting.
Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells.
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2one vector system (co-expressing spCas9and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench54) and prepared in the Yale Cancer Center Functional Genomics core). pMSCVU6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatiblewithBio-RadS3e sorting.
Plasmid Preparation:Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-Rad S3e sorting.
Article Title: The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8 + T Cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796 ; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-rad S3e sorting.
Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells.
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2one vector system (co-expressing spCas9and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench54) and prepared in the Yale Cancer Center Functional Genomics core). pMSCVU6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatiblewithBio-RadS3e sorting.
Functional Assay:Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-Rad S3e sorting.
Article Title: The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8 + T Cells
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2 one vector system (co-expressing spCas9 and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench ) and prepared in the Yale Cancer Center Functional Genomics core). pMSCV-U6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796 ; http://n2t.net/addgene:102796 ; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatible with Bio-rad S3e sorting.
Article Title: The CUL5 E3 ligase complex negatively regulates central signaling pathways in CD8 + T cells.
Article Snippet: .. The human Brie genome-wide CRISPR knockout pooled library in the pLentiCRISPRv2one vector system (co-expressing spCas9and sgRNA), with four sgRNAs per gene, was obtained from Addgene (Addgene # 73632, a gift from David Root and John Doench54) and prepared in the Yale Cancer Center Functional Genomics core). pMSCVU6sgRNA(BbsI)-PGKpuro2ABFP was a gift from Sarah Teichmann (Addgene plasmid # 102796; http://n2t.net/addgene:102796; RRID:Addgene_102796). .. PGKpuro2ABFP was replaced by EF1a-core promoter-mScarlet gblock (IDT) to generate pMSCV-guide-EF1a-mScarlet vector compatiblewithBio-RadS3e sorting.
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