pcag gfpd2 Search Results


gfp gfpd2  (Addgene inc)
93
Addgene inc gfp gfpd2
Figure 2. Comparative efficacy of 5′ DREDGE implemented with five different Cas RNases and their cognate DRs. (A) Overview of the construction of the vectors encoding <t>GFPd2</t> with individual cognate DRs (green) in the 5′ UTR (top) and vectors co-expressing different Cas RNases (red) together with mCherry (bottom). Constructs lacking a DR or RNase served as controls. (B) Cell cytometry results from MEFs transiently transfected with the constructs in (A). Depicted are the percentages of cells in Q2 relative to controls (top), which were quantified from log-log plots of GFP vs. mCherry RFU values (n = 3 replicates), with representative plots for each condition provided (bottom). (C) GFP intensity in mCherry-positive cells, normalized to controls, derived from the RFU plots shown in (B), together with representative images of GFP fluorescence in cells from the various conditions (bottom), which were acquired immediately prior to cytometry.
Gfp Gfpd2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gfp gfpd2/product/Addgene inc
Average 93 stars, based on 1 article reviews
gfp gfpd2 - by Bioz Stars, 2026-05
93/100 stars
  Buy from Supplier

Image Search Results


Figure 2. Comparative efficacy of 5′ DREDGE implemented with five different Cas RNases and their cognate DRs. (A) Overview of the construction of the vectors encoding GFPd2 with individual cognate DRs (green) in the 5′ UTR (top) and vectors co-expressing different Cas RNases (red) together with mCherry (bottom). Constructs lacking a DR or RNase served as controls. (B) Cell cytometry results from MEFs transiently transfected with the constructs in (A). Depicted are the percentages of cells in Q2 relative to controls (top), which were quantified from log-log plots of GFP vs. mCherry RFU values (n = 3 replicates), with representative plots for each condition provided (bottom). (C) GFP intensity in mCherry-positive cells, normalized to controls, derived from the RFU plots shown in (B), together with representative images of GFP fluorescence in cells from the various conditions (bottom), which were acquired immediately prior to cytometry.

Journal: Cells

Article Title: 5' DREDGE: Direct Repeat-Enabled Downregulation of Gene Expression via the 5' UTR of Target Genes.

doi: 10.3390/cells14120866

Figure Lengend Snippet: Figure 2. Comparative efficacy of 5′ DREDGE implemented with five different Cas RNases and their cognate DRs. (A) Overview of the construction of the vectors encoding GFPd2 with individual cognate DRs (green) in the 5′ UTR (top) and vectors co-expressing different Cas RNases (red) together with mCherry (bottom). Constructs lacking a DR or RNase served as controls. (B) Cell cytometry results from MEFs transiently transfected with the constructs in (A). Depicted are the percentages of cells in Q2 relative to controls (top), which were quantified from log-log plots of GFP vs. mCherry RFU values (n = 3 replicates), with representative plots for each condition provided (bottom). (C) GFP intensity in mCherry-positive cells, normalized to controls, derived from the RFU plots shown in (B), together with representative images of GFP fluorescence in cells from the various conditions (bottom), which were acquired immediately prior to cytometry.

Article Snippet: Vectors for Transient Transfection Experiments The vector expressing destabilized GFP (GFPd2) was generated by modifying Addgene plasmid #14760 [13] to include a puromycin resistance cassette, as described [4].

Techniques: Expressing, Construct, Cytometry, Transfection, Derivative Assay, Fluorescence

Figure 3. Characterization of 5′ DREDGE using an inducible system. (A) Cartoon depicting the generation of clonal cell lines constitutively expressing GFPd2 with a single Cas12a or Csy4 DR in the 5′ UTR (or no DR as a control). (B) Generation of double-stable lines for inducible Cas RNase expression. The lines in (A) were used to generate lines that also stably express either dCas12a or Csy4 (or no RNase as a control) in a Dox-regulatable manner. (C,D) Performance of the double- stable cell lines from (B), assessed using the percentage of cells in Q2 (C) and mean GFP RFU (D) in the absence or presence of Dox (1 µg/mL). Data are shown as the mean ± SEM normalized to Dox-treated No-RNase controls; n = 2–3 per condition. (E,F) Dox dose–response experiments with double-stable cell lines inducibly expressing dCas12a or Csy4. Graphs depict responsiveness to a range of concentrations of Dox, quantified in terms of the percentage mCherry-positive cells that were also GFP-positive (E) or the mean GFP RFU in all cells (F). Mean IC50 values are shown. Data are shown as the mean ± SEM for 2–3 replicates per condition, normalized to values in the absence of Dox for each line. (G) Temporal dynamics of GFPd2 expression in cell lines inducibly expressing either dCas12a or Csy4 following addition (solid lines) or withdrawal (dashed lines) of Dox (1 µg/mL). Data are shown as the mean ± SEM for 2–3 independent experiments, normalized to No-Dox controls. Mean t1/2 values are indicated. (H,I) Relative expression of GFPd2 mRNA in the absence versus the presence of Dox (1 µg/mL) in double-stable cell lines instantiating 5′ DREDGE with Cas12a (H) and Csy4 (I). Note the dramatic increase in GFPd2 mRNA levels triggered by dCas12a expression (H). Data are mean ± SEM for 3 independent experiments, expressed as a percentage of No-Dox controls.

Journal: Cells

Article Title: 5' DREDGE: Direct Repeat-Enabled Downregulation of Gene Expression via the 5' UTR of Target Genes.

doi: 10.3390/cells14120866

Figure Lengend Snippet: Figure 3. Characterization of 5′ DREDGE using an inducible system. (A) Cartoon depicting the generation of clonal cell lines constitutively expressing GFPd2 with a single Cas12a or Csy4 DR in the 5′ UTR (or no DR as a control). (B) Generation of double-stable lines for inducible Cas RNase expression. The lines in (A) were used to generate lines that also stably express either dCas12a or Csy4 (or no RNase as a control) in a Dox-regulatable manner. (C,D) Performance of the double- stable cell lines from (B), assessed using the percentage of cells in Q2 (C) and mean GFP RFU (D) in the absence or presence of Dox (1 µg/mL). Data are shown as the mean ± SEM normalized to Dox-treated No-RNase controls; n = 2–3 per condition. (E,F) Dox dose–response experiments with double-stable cell lines inducibly expressing dCas12a or Csy4. Graphs depict responsiveness to a range of concentrations of Dox, quantified in terms of the percentage mCherry-positive cells that were also GFP-positive (E) or the mean GFP RFU in all cells (F). Mean IC50 values are shown. Data are shown as the mean ± SEM for 2–3 replicates per condition, normalized to values in the absence of Dox for each line. (G) Temporal dynamics of GFPd2 expression in cell lines inducibly expressing either dCas12a or Csy4 following addition (solid lines) or withdrawal (dashed lines) of Dox (1 µg/mL). Data are shown as the mean ± SEM for 2–3 independent experiments, normalized to No-Dox controls. Mean t1/2 values are indicated. (H,I) Relative expression of GFPd2 mRNA in the absence versus the presence of Dox (1 µg/mL) in double-stable cell lines instantiating 5′ DREDGE with Cas12a (H) and Csy4 (I). Note the dramatic increase in GFPd2 mRNA levels triggered by dCas12a expression (H). Data are mean ± SEM for 3 independent experiments, expressed as a percentage of No-Dox controls.

Article Snippet: Vectors for Transient Transfection Experiments The vector expressing destabilized GFP (GFPd2) was generated by modifying Addgene plasmid #14760 [13] to include a puromycin resistance cassette, as described [4].

Techniques: Expressing, Control, Stable Transfection