Recombinant:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
Plasmid Preparation:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
Expressing:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
Luciferase:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
Amplification:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
Sequencing:Article Title: The intrinsically disordered region of eIF5B stimulates IRES usage and nucleates biological granule formation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant DNA pCRIS-PITChv2-BSD-dTAG (BRD4) James Bradner & Behnam Nabet Addgene plasmid # 91792 pX330-U6-Chimeric_BB-CBh-hSpCas9 Feng Zhang Addgene plasmid # 42230 pJDS246 expressing cas9 Keith Joung Addgene plasmid # 43861 pGL3 Promega Empty firefly luciferase vector RC203227 Origene DENR (Myc-DDK-tagged)-Human density-regulated protein (DENR) expression vector RC204303 Origene EIF2A (Myc-DDK-tagged)-Human eukaryotic translation initiation factor 2A, 65kDa (EIF2A) expression vector RC200224 Origene MCTS1 (Myc-DDK-tagged)-Human malignant T cell amplified sequence 1 (MCTS1) expression vector MR208977 Origene Eif2d (Myc-DDK-tagged) - Mouse eukaryotic translation initiation factor 2D (Eif2d) expression vector 5496504 Transomics MGC expression clone 5496504 (EIF5B) pGLRSVRF Olson et al.26 Bicistronic vector p91792_eIF5B_BLAST This study EIF5B targeting vector p91792_eIF5B_NEO This study EIF5B targeting vector phU6_eIF5B_gRNA_2654 This study EIF5B gRNA vector pGLRSVRF_INR Olson et al.26 Bicistronic vector pGLRSVRF_IGFR Olson et al.26 Bicistronic vector pGLRSVRF_HCV Olson et al.26 Bicistronic vector pGLRSVRF_beta globin This study Bicistronic vector pCMV_INR_luc_spinach3 This study Spinach tagged reporter pCMV_BG_luc_spinach3 This study Spinach tagged reporter pCMV_EIF5B_NTR This study EIF5B fragment expression vectors pCMV_EIF5B_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NS_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NL_CTR This study EIF5B fragment expression vectors pCMV_EIF5B_NM_CTR This study EIF5B fragment expression vectors pCMV_mNG-eIF5B This study EIF5B fusion to mNG pCMV_mNG-NTR This study EIF5B fusion to mNG pCMV_mNG-CTR This study EIF5B fusion to mNG pCMV_mRuby-eIF5B This study EIF5B fusion to mRuby2 mNeonGreen-mRuby2-FRET-10 Michael Davidson Addgene plasmid #58179 pHTN Promega Halo tag vector pHTN_EIF5B This study Expression plasmid for halo- EIF5B pHTN_NTR This study Expression plasmid for halo- NTR pET28_EIF5B_CTR This study Expression plasmid for EIF5B CTR pCMV-15F11-HA-mEGFP Tim Stasevich Addgene plasmid #129590 pCMV_Frankenberry This study mRUby fusion to 2 3 15F11 pN1/G3BP1-iRFP Yasushi Okada Addgene #129339 pCMV G3BP1 mNG This study G3BP1 fusion to mNG pCMV G3BP1 mRuby This study G3BP1 fusion to mRuby2 (Continued on next page) Cell Reports 42, 113283, October 31, 2023 15
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