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Addgene inc gfp only control vector
dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of <t>GFP</t> +ve cells after infection with no virus (green), <t>hTERT</t> with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)
Gfp Only Control Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gfp only control vector/product/Addgene inc
Average 91 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gfp only control vector - by Bioz Stars, 2020-08
91/100 stars

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1) Product Images from "Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors"

Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors

Journal: Nature Communications

doi: 10.1038/s41467-017-01078-2

dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of GFP +ve cells after infection with no virus (green), hTERT with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)
Figure Legend Snippet: dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of GFP +ve cells after infection with no virus (green), hTERT with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)

Techniques Used: Colorimetric Assay, Transfection, Light Microscopy, Staining, Infection, Marker

Related Articles

Marker:

Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
Article Snippet: .. A retroviral vector expressing hTERT with a GFP marker (Addgene #69809) and a GFP-only control vector (Addgene #21654) were transfected into the Plat-A cells using jetprime. .. Fresh media without selection antibiotics was put on Plat-A cells 24 h after transfection.

Transfection:

Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
Article Snippet: .. A retroviral vector expressing hTERT with a GFP marker (Addgene #69809) and a GFP-only control vector (Addgene #21654) were transfected into the Plat-A cells using jetprime. .. Fresh media without selection antibiotics was put on Plat-A cells 24 h after transfection.

Plasmid Preparation:

Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
Article Snippet: .. A retroviral vector expressing hTERT with a GFP marker (Addgene #69809) and a GFP-only control vector (Addgene #21654) were transfected into the Plat-A cells using jetprime. .. Fresh media without selection antibiotics was put on Plat-A cells 24 h after transfection.

Expressing:

Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
Article Snippet: .. A retroviral vector expressing hTERT with a GFP marker (Addgene #69809) and a GFP-only control vector (Addgene #21654) were transfected into the Plat-A cells using jetprime. .. Fresh media without selection antibiotics was put on Plat-A cells 24 h after transfection.

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  • 91
    Addgene inc gfp only control vector
    dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of <t>GFP</t> +ve cells after infection with no virus (green), <t>hTERT</t> with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)
    Gfp Only Control Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp only control vector/product/Addgene inc
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gfp only control vector - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    85
    Addgene inc control gfp only virus
    dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of <t>GFP</t> +ve cells after infection with no virus (green), <t>hTERT</t> with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)
    Control Gfp Only Virus, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/control gfp only virus/product/Addgene inc
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    control gfp only virus - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

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    dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of GFP +ve cells after infection with no virus (green), hTERT with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)

    Journal: Nature Communications

    Article Title: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors

    doi: 10.1038/s41467-017-01078-2

    Figure Lengend Snippet: dCas9 3A3L or dCas9 3A targeting prevents senescence and increases proliferation. a , b Proliferation was assessed using a colorimetric assay 10, 15 and 20 days after targeting dCas9 3A3L to HIC1 , RASSF1 , PTEN and CDKN2A and magnetic sorting in primary myoepithelial cells from donors 1, 2 and 3. Dotted line represents the average absorbance from three dCas9 3A3LΔ targeted wells at each timepoint. The graphs show the average difference in absorbance as fold change compared to 3A3LΔ average (mean ± SEM, n = 3). c Cumulative population doublings over time for untransfected primary myoepithelial cells and those transfected with dCas9 3A3L or 3A3L∆ targeting the four genes without magnetic sorting from donor one. Fifty thousand cells were seeded at the start of each passage and cells counted after 5 days (mean ± SEM, n = 3, where error bars are smaller than points plotted they are not shown). ( d ) Light microscopy images of β-gal staining from untransfected (top left) primary myoepithelial cells from donor 1 and those transfected with dCas9 3A3L (bottom two) or 3A3L∆ (top right) targeting the four genes without magnetic sorting 20 days after transfection, blue/green colouring shows β-gal +ve cells; scale bar represents 125 µm. e Proliferation was assessed using a colorimetric assay 15, 20 and 25 days after targeting the four genes with dCas9 3A3L (red), dCas9 3A (orange), dCas9 3A-C706A-3L (yellow), dCas9 3A3L∆ (green) or dCas9m4 (pale green), or targeting only dCas9 3A3L without gRNAs (blue) without magnetic sorting using donor 1 myoepithelial cells. The data is shown as fold change compared to the average absorbance from 3 wells without cells (mean ± SEM, n = 3). f The percentage of GFP +ve cells after infection with no virus (green), hTERT with GFP marker (pink) or GFP control (purple). At infection day 0 cells are 38 days post-transfection with dCas9 3A3L targeting the four genes (mean ± SEM; n = 3, error bars are not shown where smaller than points plotted)

    Article Snippet: A retroviral vector expressing hTERT with a GFP marker (Addgene #69809) and a GFP-only control vector (Addgene #21654) were transfected into the Plat-A cells using jetprime.

    Techniques: Colorimetric Assay, Transfection, Light Microscopy, Staining, Infection, Marker