rtetr cbx7 deletion Search Results


rtetr cbx7 deletion  (Addgene inc)
92
Addgene inc rtetr cbx7 deletion
Fig. 1. Newly established Polycomb silencing near developmental genes, but not at ubiquitously expressed genes, is heritable. (A) Schematic diagram illustrating the strategy for inducible Polycomb silencing by expression of <t>rTetR-CBX7</t> as a fusion with mCherry-2A in cells carrying the 5xtetO-CITRINE reporter at various develop- mental and ubiquitously expressed genes. See figs. S1 and S2 for exact coordinates of reporter insertion. (B) Representative fluorescence images showing mCherry and CITRINE expression before and after establishment of silencing (−Dox and +Dox, respectively). (C) Flow cytometry histograms showing CITRINE expression before and after establishment of silencing with doxycycline addition (−Dox and +Dox) and at the indicated days after removal of doxycycline (Dox removal) at developmental genes WT1, HOXD, and EN2. Percentages (%) indicate the fraction of CITRINE negative cells. (D) Same as (C) but with the reporter inserted near ubiquitously expressed genes B2M and TFRC.
Rtetr Cbx7 Deletion, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rtetr+cbx7+deletion/pm38718120-245-26-33?v=Addgene+inc
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eadl4529 2024 8 may  (Addgene inc)
92
Addgene inc eadl4529 2024 8 may
Fig. 1. Newly established Polycomb silencing near developmental genes, but not at ubiquitously expressed genes, is heritable. (A) Schematic diagram illustrating the strategy for inducible Polycomb silencing by expression of <t>rTetR-CBX7</t> as a fusion with mCherry-2A in cells carrying the 5xtetO-CITRINE reporter at various develop- mental and ubiquitously expressed genes. See figs. S1 and S2 for exact coordinates of reporter insertion. (B) Representative fluorescence images showing mCherry and CITRINE expression before and after establishment of silencing (−Dox and +Dox, respectively). (C) Flow cytometry histograms showing CITRINE expression before and after establishment of silencing with doxycycline addition (−Dox and +Dox) and at the indicated days after removal of doxycycline (Dox removal) at developmental genes WT1, HOXD, and EN2. Percentages (%) indicate the fraction of CITRINE negative cells. (D) Same as (C) but with the reporter inserted near ubiquitously expressed genes B2M and TFRC.
Eadl4529 2024 8 May, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rtetr+cbx7+deletion/pm38718120-245-1-10?v=Addgene+inc
Average 92 stars, based on 1 article reviews
eadl4529 2024 8 may - by Bioz Stars, 2026-08
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pspcas9 bb 2a puro  (Addgene inc)
96
Addgene inc pspcas9 bb 2a puro
Fig. 1. Newly established Polycomb silencing near developmental genes, but not at ubiquitously expressed genes, is heritable. (A) Schematic diagram illustrating the strategy for inducible Polycomb silencing by expression of <t>rTetR-CBX7</t> as a fusion with mCherry-2A in cells carrying the 5xtetO-CITRINE reporter at various develop- mental and ubiquitously expressed genes. See figs. S1 and S2 for exact coordinates of reporter insertion. (B) Representative fluorescence images showing mCherry and CITRINE expression before and after establishment of silencing (−Dox and +Dox, respectively). (C) Flow cytometry histograms showing CITRINE expression before and after establishment of silencing with doxycycline addition (−Dox and +Dox) and at the indicated days after removal of doxycycline (Dox removal) at developmental genes WT1, HOXD, and EN2. Percentages (%) indicate the fraction of CITRINE negative cells. (D) Same as (C) but with the reporter inserted near ubiquitously expressed genes B2M and TFRC.
Pspcas9 Bb 2a Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rtetr+cbx7+deletion/bio_rxiv__2023__02__27__530239-187-77-80?v=Addgene+inc
Average 96 stars, based on 1 article reviews
pspcas9 bb 2a puro - by Bioz Stars, 2026-08
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Image Search Results


Fig. 1. Newly established Polycomb silencing near developmental genes, but not at ubiquitously expressed genes, is heritable. (A) Schematic diagram illustrating the strategy for inducible Polycomb silencing by expression of rTetR-CBX7 as a fusion with mCherry-2A in cells carrying the 5xtetO-CITRINE reporter at various develop- mental and ubiquitously expressed genes. See figs. S1 and S2 for exact coordinates of reporter insertion. (B) Representative fluorescence images showing mCherry and CITRINE expression before and after establishment of silencing (−Dox and +Dox, respectively). (C) Flow cytometry histograms showing CITRINE expression before and after establishment of silencing with doxycycline addition (−Dox and +Dox) and at the indicated days after removal of doxycycline (Dox removal) at developmental genes WT1, HOXD, and EN2. Percentages (%) indicate the fraction of CITRINE negative cells. (D) Same as (C) but with the reporter inserted near ubiquitously expressed genes B2M and TFRC.

Journal: Science advances

Article Title: Genomic context- and H2AK119 ubiquitination-dependent inheritance of human Polycomb silencing.

doi: 10.1126/sciadv.adl4529

Figure Lengend Snippet: Fig. 1. Newly established Polycomb silencing near developmental genes, but not at ubiquitously expressed genes, is heritable. (A) Schematic diagram illustrating the strategy for inducible Polycomb silencing by expression of rTetR-CBX7 as a fusion with mCherry-2A in cells carrying the 5xtetO-CITRINE reporter at various develop- mental and ubiquitously expressed genes. See figs. S1 and S2 for exact coordinates of reporter insertion. (B) Representative fluorescence images showing mCherry and CITRINE expression before and after establishment of silencing (−Dox and +Dox, respectively). (C) Flow cytometry histograms showing CITRINE expression before and after establishment of silencing with doxycycline addition (−Dox and +Dox) and at the indicated days after removal of doxycycline (Dox removal) at developmental genes WT1, HOXD, and EN2. Percentages (%) indicate the fraction of CITRINE negative cells. (D) Same as (C) but with the reporter inserted near ubiquitously expressed genes B2M and TFRC.

Article Snippet: 10, eadl4529 (2024) 8 May 2024 12 of 15 V2.0 (Addgene, plasmid no. 62988) and transfected into HEK293FT using Lipofectamine 2000 (Thermo Fisher Scientific,). sgRNAs for rTetR- CBX7 deletion were cloned into LentiCRISPRv2 (Addgene, 52961).

Techniques: Expressing, Fluorescence, Flow Cytometry

Fig. 6. Interactions between Polycomb complexes. (A) Volcano plot displaying the results of MS identification of proteins enriched in Flag immunoprecipitations from cells expressing rTetR-CBX7-3xFlag relative to untagged cells from two independent experiments. Subunits of the cPRC1, vPRC1, PRC2, rixosome, and histones are high- lighted. (B) Coimmunoprecipitations showing the association of RING1B with PRC1 subunit CBX7 and PRC2 subunits EZH2 in HEK 293FT cells. IB, immunoblot. (C) Coim- munoprecipitations showing the association of CBX7 and PRC2 subunits EZH2 in HEK 293FT cells. (D) Pulldown assays using bead-immobilized GST or GST-BCORL(S1474-G1644) proteins and Strep-RBBP7, showing binding of Strep-RBBP7 to the GST-BCORL fragment.

Journal: Science advances

Article Title: Genomic context- and H2AK119 ubiquitination-dependent inheritance of human Polycomb silencing.

doi: 10.1126/sciadv.adl4529

Figure Lengend Snippet: Fig. 6. Interactions between Polycomb complexes. (A) Volcano plot displaying the results of MS identification of proteins enriched in Flag immunoprecipitations from cells expressing rTetR-CBX7-3xFlag relative to untagged cells from two independent experiments. Subunits of the cPRC1, vPRC1, PRC2, rixosome, and histones are high- lighted. (B) Coimmunoprecipitations showing the association of RING1B with PRC1 subunit CBX7 and PRC2 subunits EZH2 in HEK 293FT cells. IB, immunoblot. (C) Coim- munoprecipitations showing the association of CBX7 and PRC2 subunits EZH2 in HEK 293FT cells. (D) Pulldown assays using bead-immobilized GST or GST-BCORL(S1474-G1644) proteins and Strep-RBBP7, showing binding of Strep-RBBP7 to the GST-BCORL fragment.

Article Snippet: 10, eadl4529 (2024) 8 May 2024 12 of 15 V2.0 (Addgene, plasmid no. 62988) and transfected into HEK293FT using Lipofectamine 2000 (Thermo Fisher Scientific,). sgRNAs for rTetR- CBX7 deletion were cloned into LentiCRISPRv2 (Addgene, 52961).

Techniques: Expressing, Western Blot, Binding Assay

Fig. 7. Interactions involving PRCs and histone modifications that mediate heritable silencing. (A) Diagram showing the expected rTetR-CBX7-initiated re- cruitment of PRCs (cPRC1, vPRC1, and PRC2) leading to H2AK119 mono- ubiquitination (H2AK119ub1) and H3K27 trimethylation (H3K27me3). (B) Summary of interactions in wild-type cells at a developmental gene targeted by Polycomb. (C) In the absence of H3K27me3 in EED-3A cells, intact PRC2 and PRC1 complexes mediate heritable silencing. (D) At housekeeping genes, Polycomb silencing is not heritable due to the absence of extended CpG islands (eCGI) and/or other differ- ences between housekeeping genes and developmental genes.

Journal: Science advances

Article Title: Genomic context- and H2AK119 ubiquitination-dependent inheritance of human Polycomb silencing.

doi: 10.1126/sciadv.adl4529

Figure Lengend Snippet: Fig. 7. Interactions involving PRCs and histone modifications that mediate heritable silencing. (A) Diagram showing the expected rTetR-CBX7-initiated re- cruitment of PRCs (cPRC1, vPRC1, and PRC2) leading to H2AK119 mono- ubiquitination (H2AK119ub1) and H3K27 trimethylation (H3K27me3). (B) Summary of interactions in wild-type cells at a developmental gene targeted by Polycomb. (C) In the absence of H3K27me3 in EED-3A cells, intact PRC2 and PRC1 complexes mediate heritable silencing. (D) At housekeeping genes, Polycomb silencing is not heritable due to the absence of extended CpG islands (eCGI) and/or other differ- ences between housekeeping genes and developmental genes.

Article Snippet: 10, eadl4529 (2024) 8 May 2024 12 of 15 V2.0 (Addgene, plasmid no. 62988) and transfected into HEK293FT using Lipofectamine 2000 (Thermo Fisher Scientific,). sgRNAs for rTetR- CBX7 deletion were cloned into LentiCRISPRv2 (Addgene, 52961).

Techniques: Ubiquitin Proteomics