Generated:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Article Title: Splicing-guaranteed artificial introns enable efficient homology-directed repair at internal exons and streamlined cell engineering
Article Snippet: .. DSBs were generated by CRISPR–Cas9 technology with LentiCRISPR-V2 (Addgene, #98290) carrying the indicated intron-targeting sgRNAs (RELA: 5’-UCA CUG AGC UCC CGG UCG GA-3’, EGFR: 5’-GCU GGC CAA ACU GCU GGG UG-3’, PIK3CA: 5’-AUG CAU GCU GUU UAA UUG UG-3’, EMX1: 5’-GAG UCC GAG CAG AAG AAG AA-3’, FANCF: 5’-GGA AUC CCU UCU GCA GCA CC-3’). ..
Expressing:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Article Title: The spindle-associated antisense RNA ARHGEF17-AS1 functionally contributes to mitotic spindle integrity.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides siRNAs and LNA antisense oligonucleotides used in this study Sigma-Aldrich; Qiagen; this paper See Table S1 qPCR and ChIRP-qPCR primers used in this study This paper See Table S2 sgRNA and shRNA oligonucleotides used in this study Sigma-Aldrich; this paper See Table S3 ChIRP and ChOP capture probes used in this study Sigma-Aldrich; this paper See Table S4 ARHGEF17-AS1 knockout sequencing primers This paper See Table S5 Custom RNAscope probe set targeting human ARHGEF17-AS1 This paper/Advanced Cell Diagnostics/Bio-Techne Custom Probe design targeting second and third exon of AS1. .. Recombinant DNA pcDNA3.1+ mammalian expression vector Invitrogen Cat# V79020 pcDNA3.1-ARHGEF17-AS1 This paper N/A pLKO-Tet-On-shRNA-Control Junqueira Alves et al.29; Addgene Addgene plasmid #98398; RRID: Addgene_98398 pLKO.1-ARHGEF17-AS1-sh1 This paper N/A pLKO.1-ARHGEF17-AS1-sh2 This paper N/A pLVX-TetOne-Puro vector Takara Bio Cat# 631847 pLVX-TetOne-ARHGEF17-AS1 This paper N/A pLVX-TetOne-LacZ This paper N/A pLenti_Empty (EVC_Flag) Gift from Thomas Tuschl N/A IC74-mfGFP cytoplasmic dynein intermediate-chain expression plasmid Gift from Takashi Murayama; Kobayashi and Murayama30 N/A pSpCas9(BB)-2A-GFP (PX458) Ran et al.31; Addgene Addgene plasmid #48138; RRID: Addgene_48138 pSpCas9(BB)-2A-GFP (PX458)-sg1-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg1ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg3ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro Stringer et al.32; Addgene Addgene plasmid #98290; RRID: Addgene_98290 lentiCRISPRv2 puro-LacZ This paper N/A lentiCRISPRv2 puro-sg1-ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro-sg2-ARHGEF17-AS1 This paper N/A pLENTI_RPS3 Meyer et al.33; Addgene Addgene plasmid #127128; RRID: Addgene_127128 pLENTI_RPS3_K214R Meyer et al.33; Addgene Addgene plasmid #127132; RRID: Addgene_127132 pLENTI_RPS3_K227R Meyer et al.33; Addgene Addgene plasmid #127133; RRID: Addgene_127133 pLENTI_RPS3_K230R Meyer et al.33; Addgene Addgene plasmid #127134; RRID: Addgene_127134 scramble shRNA Sarbassov et al.34; Addgene Addgene plasmid #1864; RRID: Addgene_1864 (Continued on next page) 18 Cell Reports 45, 118023, October 27, 2026 .. HeLa (Homo sapiens; cervical adenocarcinoma; 31-year-old female donor; ATCC CCL-2) and A549 (Homo sapiens; lung adenocarcinoma; 58-year-old male donor; ECACC 86012804) cells were obtained from ATCC and The European Collection of Authenticated Cell Cultures (ECACC), respectively.
Control:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Single Cell:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Cloning:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Biomarker Discovery:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Western Blot:Article Title: Antagonistic roles of histone H3 lysine methyltransferases dictate tumor immune surveillance
Article Snippet: .. KP-Nsd2KO cell lines were generated by transducing LG1233 cells with lentiCRISPRv2Puro (Addgene 98290) expressing an sgRNA targeting Nsd2 or βgal (as a control), followed by single-cell cloning (described above), and validation of NSD2 loss by immunoblotting. .. KP-H3K36MSetd2KO-EZH2KO cell lines were produced using lentiCRISPRv2mCherry (Addgene 99154) with sgRNAs targeting either EZH2 or βgal (as a control) and transducing KP-H3K36M-Setd2KO cells with these constructs.
Recombinant:Article Title: The spindle-associated antisense RNA ARHGEF17-AS1 functionally contributes to mitotic spindle integrity.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides siRNAs and LNA antisense oligonucleotides used in this study Sigma-Aldrich; Qiagen; this paper See Table S1 qPCR and ChIRP-qPCR primers used in this study This paper See Table S2 sgRNA and shRNA oligonucleotides used in this study Sigma-Aldrich; this paper See Table S3 ChIRP and ChOP capture probes used in this study Sigma-Aldrich; this paper See Table S4 ARHGEF17-AS1 knockout sequencing primers This paper See Table S5 Custom RNAscope probe set targeting human ARHGEF17-AS1 This paper/Advanced Cell Diagnostics/Bio-Techne Custom Probe design targeting second and third exon of AS1. .. Recombinant DNA pcDNA3.1+ mammalian expression vector Invitrogen Cat# V79020 pcDNA3.1-ARHGEF17-AS1 This paper N/A pLKO-Tet-On-shRNA-Control Junqueira Alves et al.29; Addgene Addgene plasmid #98398; RRID: Addgene_98398 pLKO.1-ARHGEF17-AS1-sh1 This paper N/A pLKO.1-ARHGEF17-AS1-sh2 This paper N/A pLVX-TetOne-Puro vector Takara Bio Cat# 631847 pLVX-TetOne-ARHGEF17-AS1 This paper N/A pLVX-TetOne-LacZ This paper N/A pLenti_Empty (EVC_Flag) Gift from Thomas Tuschl N/A IC74-mfGFP cytoplasmic dynein intermediate-chain expression plasmid Gift from Takashi Murayama; Kobayashi and Murayama30 N/A pSpCas9(BB)-2A-GFP (PX458) Ran et al.31; Addgene Addgene plasmid #48138; RRID: Addgene_48138 pSpCas9(BB)-2A-GFP (PX458)-sg1-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg1ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg3ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro Stringer et al.32; Addgene Addgene plasmid #98290; RRID: Addgene_98290 lentiCRISPRv2 puro-LacZ This paper N/A lentiCRISPRv2 puro-sg1-ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro-sg2-ARHGEF17-AS1 This paper N/A pLENTI_RPS3 Meyer et al.33; Addgene Addgene plasmid #127128; RRID: Addgene_127128 pLENTI_RPS3_K214R Meyer et al.33; Addgene Addgene plasmid #127132; RRID: Addgene_127132 pLENTI_RPS3_K227R Meyer et al.33; Addgene Addgene plasmid #127133; RRID: Addgene_127133 pLENTI_RPS3_K230R Meyer et al.33; Addgene Addgene plasmid #127134; RRID: Addgene_127134 scramble shRNA Sarbassov et al.34; Addgene Addgene plasmid #1864; RRID: Addgene_1864 (Continued on next page) 18 Cell Reports 45, 118023, October 27, 2026 .. HeLa (Homo sapiens; cervical adenocarcinoma; 31-year-old female donor; ATCC CCL-2) and A549 (Homo sapiens; lung adenocarcinoma; 58-year-old male donor; ECACC 86012804) cells were obtained from ATCC and The European Collection of Authenticated Cell Cultures (ECACC), respectively.
Plasmid Preparation:Article Title: The spindle-associated antisense RNA ARHGEF17-AS1 functionally contributes to mitotic spindle integrity.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides siRNAs and LNA antisense oligonucleotides used in this study Sigma-Aldrich; Qiagen; this paper See Table S1 qPCR and ChIRP-qPCR primers used in this study This paper See Table S2 sgRNA and shRNA oligonucleotides used in this study Sigma-Aldrich; this paper See Table S3 ChIRP and ChOP capture probes used in this study Sigma-Aldrich; this paper See Table S4 ARHGEF17-AS1 knockout sequencing primers This paper See Table S5 Custom RNAscope probe set targeting human ARHGEF17-AS1 This paper/Advanced Cell Diagnostics/Bio-Techne Custom Probe design targeting second and third exon of AS1. .. Recombinant DNA pcDNA3.1+ mammalian expression vector Invitrogen Cat# V79020 pcDNA3.1-ARHGEF17-AS1 This paper N/A pLKO-Tet-On-shRNA-Control Junqueira Alves et al.29; Addgene Addgene plasmid #98398; RRID: Addgene_98398 pLKO.1-ARHGEF17-AS1-sh1 This paper N/A pLKO.1-ARHGEF17-AS1-sh2 This paper N/A pLVX-TetOne-Puro vector Takara Bio Cat# 631847 pLVX-TetOne-ARHGEF17-AS1 This paper N/A pLVX-TetOne-LacZ This paper N/A pLenti_Empty (EVC_Flag) Gift from Thomas Tuschl N/A IC74-mfGFP cytoplasmic dynein intermediate-chain expression plasmid Gift from Takashi Murayama; Kobayashi and Murayama30 N/A pSpCas9(BB)-2A-GFP (PX458) Ran et al.31; Addgene Addgene plasmid #48138; RRID: Addgene_48138 pSpCas9(BB)-2A-GFP (PX458)-sg1-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg1ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg3ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro Stringer et al.32; Addgene Addgene plasmid #98290; RRID: Addgene_98290 lentiCRISPRv2 puro-LacZ This paper N/A lentiCRISPRv2 puro-sg1-ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro-sg2-ARHGEF17-AS1 This paper N/A pLENTI_RPS3 Meyer et al.33; Addgene Addgene plasmid #127128; RRID: Addgene_127128 pLENTI_RPS3_K214R Meyer et al.33; Addgene Addgene plasmid #127132; RRID: Addgene_127132 pLENTI_RPS3_K227R Meyer et al.33; Addgene Addgene plasmid #127133; RRID: Addgene_127133 pLENTI_RPS3_K230R Meyer et al.33; Addgene Addgene plasmid #127134; RRID: Addgene_127134 scramble shRNA Sarbassov et al.34; Addgene Addgene plasmid #1864; RRID: Addgene_1864 (Continued on next page) 18 Cell Reports 45, 118023, October 27, 2026 .. HeLa (Homo sapiens; cervical adenocarcinoma; 31-year-old female donor; ATCC CCL-2) and A549 (Homo sapiens; lung adenocarcinoma; 58-year-old male donor; ECACC 86012804) cells were obtained from ATCC and The European Collection of Authenticated Cell Cultures (ECACC), respectively.
shRNA:Article Title: The spindle-associated antisense RNA ARHGEF17-AS1 functionally contributes to mitotic spindle integrity.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides siRNAs and LNA antisense oligonucleotides used in this study Sigma-Aldrich; Qiagen; this paper See Table S1 qPCR and ChIRP-qPCR primers used in this study This paper See Table S2 sgRNA and shRNA oligonucleotides used in this study Sigma-Aldrich; this paper See Table S3 ChIRP and ChOP capture probes used in this study Sigma-Aldrich; this paper See Table S4 ARHGEF17-AS1 knockout sequencing primers This paper See Table S5 Custom RNAscope probe set targeting human ARHGEF17-AS1 This paper/Advanced Cell Diagnostics/Bio-Techne Custom Probe design targeting second and third exon of AS1. .. Recombinant DNA pcDNA3.1+ mammalian expression vector Invitrogen Cat# V79020 pcDNA3.1-ARHGEF17-AS1 This paper N/A pLKO-Tet-On-shRNA-Control Junqueira Alves et al.29; Addgene Addgene plasmid #98398; RRID: Addgene_98398 pLKO.1-ARHGEF17-AS1-sh1 This paper N/A pLKO.1-ARHGEF17-AS1-sh2 This paper N/A pLVX-TetOne-Puro vector Takara Bio Cat# 631847 pLVX-TetOne-ARHGEF17-AS1 This paper N/A pLVX-TetOne-LacZ This paper N/A pLenti_Empty (EVC_Flag) Gift from Thomas Tuschl N/A IC74-mfGFP cytoplasmic dynein intermediate-chain expression plasmid Gift from Takashi Murayama; Kobayashi and Murayama30 N/A pSpCas9(BB)-2A-GFP (PX458) Ran et al.31; Addgene Addgene plasmid #48138; RRID: Addgene_48138 pSpCas9(BB)-2A-GFP (PX458)-sg1-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2-LacZ This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg1ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg2ARHGEF17-AS1 This paper N/A pSpCas9(BB)-2A-GFP (PX458)-sg3ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro Stringer et al.32; Addgene Addgene plasmid #98290; RRID: Addgene_98290 lentiCRISPRv2 puro-LacZ This paper N/A lentiCRISPRv2 puro-sg1-ARHGEF17-AS1 This paper N/A lentiCRISPRv2 puro-sg2-ARHGEF17-AS1 This paper N/A pLENTI_RPS3 Meyer et al.33; Addgene Addgene plasmid #127128; RRID: Addgene_127128 pLENTI_RPS3_K214R Meyer et al.33; Addgene Addgene plasmid #127132; RRID: Addgene_127132 pLENTI_RPS3_K227R Meyer et al.33; Addgene Addgene plasmid #127133; RRID: Addgene_127133 pLENTI_RPS3_K230R Meyer et al.33; Addgene Addgene plasmid #127134; RRID: Addgene_127134 scramble shRNA Sarbassov et al.34; Addgene Addgene plasmid #1864; RRID: Addgene_1864 (Continued on next page) 18 Cell Reports 45, 118023, October 27, 2026 .. HeLa (Homo sapiens; cervical adenocarcinoma; 31-year-old female donor; ATCC CCL-2) and A549 (Homo sapiens; lung adenocarcinoma; 58-year-old male donor; ECACC 86012804) cells were obtained from ATCC and The European Collection of Authenticated Cell Cultures (ECACC), respectively.
CRISPR:Article Title: Splicing-guaranteed artificial introns enable efficient homology-directed repair at internal exons and streamlined cell engineering
Article Snippet: .. DSBs were generated by CRISPR–Cas9 technology with LentiCRISPR-V2 (Addgene, #98290) carrying the indicated intron-targeting sgRNAs (RELA: 5’-UCA CUG AGC UCC CGG UCG GA-3’, EGFR: 5’-GCU GGC CAA ACU GCU GGG UG-3’, PIK3CA: 5’-AUG CAU GCU GUU UAA UUG UG-3’, EMX1: 5’-GAG UCC GAG CAG AAG AAG AA-3’, FANCF: 5’-GGA AUC CCU UCU GCA GCA CC-3’). ..
Cellular Antioxidant Activity Assay:Article Title: Splicing-guaranteed artificial introns enable efficient homology-directed repair at internal exons and streamlined cell engineering
Article Snippet: .. DSBs were generated by CRISPR–Cas9 technology with LentiCRISPR-V2 (Addgene, #98290) carrying the indicated intron-targeting sgRNAs (RELA: 5’-UCA CUG AGC UCC CGG UCG GA-3’, EGFR: 5’-GCU GGC CAA ACU GCU GGG UG-3’, PIK3CA: 5’-AUG CAU GCU GUU UAA UUG UG-3’, EMX1: 5’-GAG UCC GAG CAG AAG AAG AA-3’, FANCF: 5’-GGA AUC CCU UCU GCA GCA CC-3’). ..
Clone Assay:
Sequencing:
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