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addgene provided pcdna3 mruby2  (Addgene inc)


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    Structured Review

    Addgene inc addgene provided pcdna3 mruby2
    Addgene Provided Pcdna3 Mruby2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna3+mruby2/pcDNA3-mRuby2+(Plasmid+%2340260)/bio_rxiv__64898__2026__03__11__711183-172-0-0
    Average 93 stars, based on 80 article reviews
    addgene provided pcdna3 mruby2 - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Convergent activation of Ca 2+ permeability in two-pore channel 2 through distinct molecular routes
    Article Snippet: .. The coding sequences incorporating a 3' (Gly-Ser-Ala)3 flexible linker were subcloned into pcDNA3-mRuby2 (Addgene plasmid #40260) ( 42 ) by homologous recombination using the primer sequences listed in table S1 . .. All gene synthesis and subcloning were performed by GenScript (Piscataway, NJ).

    Article Title: Three-component CRISPR/Cas complex system and uses thereof
    Article Snippet: A fusion PCR of 5′ fragment of NLSPUFa with 3′ fragment of NLSPUb was used to create pAC1358:pmax-NLSPUFc_VP64. p65HSF1 activator ORF was amplified from MS2-P65-HSF1_GFP (Addgene: 61423) with FseI PacI sites to replace VP64 fragment in pAC164 to create pAC1410:pmax-dCas9_p65HSF1, and replace VP64 in pAC1355 and pAC1358 to create pAC1393: pmax-NLSPUFa_p65HSF1 and pAC1411:pmax-NLSPUFc_p65HSF1, respectively. .. Clover and mRuby2 were amplified from pcDNA3-Clover (Addgene #40259) and pcDNA3-mRuby2 (Addgene #40260) respectively with primers containing SgrAI and FseI cloning site, ligated with various FseI-PUF-PacI amplified from the above pAC1356 ̃1358 and vector digested from pAC149:pCR8-dCas9VP160 (Addgene #48221) to create gateway donor vectors pAC1402, pAC1403 and pAC1404 containing ORFs of Clover_PUFa and Clover_PUFc, mRuby2_PUFa, respectively. .. These ORFs are then transferred to PB3-neo vector by recombining with pAC1119:PB3-neo(−)-pmaxDEST(+) by LR Clonase (Invitrogen) to create expression vectors pAC1360 (Clover_PUFa), pAC1381 (Clover_PUFc) and pAC1362 (mRuby2_PUFa).

    Article Title: Exosomes as Conduits: Facilitating Hepatitis B Virus-Independent Hepatitis D Virus Transmission and Propagation in Hepatocytes
    Article Snippet: .. The plasmids pCIHD24 and pCIHD27 were used for the expression of S-HDAg and L-HDAg, respectively, as previously described [ , ]. pCIneo and peGFP were purchased from Promega and Clontech, respectively. pcDNA3 mRuby2 (named in pmRuby2) plasmid is from Addgene [ ]. .. Supernatants from transfected cells cultured in DMEM supplemented with 5% exosome-depleted serum were collected and clarified.

    Article Title: Tissue‐like environments shape functional interactions of HIV ‐1 with immature dendritic cells
    Article Snippet: .. The plasmid pmRuby2.Vpr, encoding Vpr fused to mRuby2, was generated by excision of the eGFP coding region of peGFP.Vpr (McDonald et al , , a kind gift of Tom Hope) using AgeI and BsrGI restriction enzymes, followed by insertion of a PCR fragment encoding mRuby2 (Lam et al , ); amplified from pcDNA3‐mRuby2, a gift from Michael Lin (Addgene plasmid # 40260) and flanked by the same restriction sites. ..

    Homologous Recombination:

    Article Title: Convergent activation of Ca 2+ permeability in two-pore channel 2 through distinct molecular routes
    Article Snippet: .. The coding sequences incorporating a 3' (Gly-Ser-Ala)3 flexible linker were subcloned into pcDNA3-mRuby2 (Addgene plasmid #40260) ( 42 ) by homologous recombination using the primer sequences listed in table S1 . .. All gene synthesis and subcloning were performed by GenScript (Piscataway, NJ).

    Amplification:

    Article Title: Three-component CRISPR/Cas complex system and uses thereof
    Article Snippet: A fusion PCR of 5′ fragment of NLSPUFa with 3′ fragment of NLSPUb was used to create pAC1358:pmax-NLSPUFc_VP64. p65HSF1 activator ORF was amplified from MS2-P65-HSF1_GFP (Addgene: 61423) with FseI PacI sites to replace VP64 fragment in pAC164 to create pAC1410:pmax-dCas9_p65HSF1, and replace VP64 in pAC1355 and pAC1358 to create pAC1393: pmax-NLSPUFa_p65HSF1 and pAC1411:pmax-NLSPUFc_p65HSF1, respectively. .. Clover and mRuby2 were amplified from pcDNA3-Clover (Addgene #40259) and pcDNA3-mRuby2 (Addgene #40260) respectively with primers containing SgrAI and FseI cloning site, ligated with various FseI-PUF-PacI amplified from the above pAC1356 ̃1358 and vector digested from pAC149:pCR8-dCas9VP160 (Addgene #48221) to create gateway donor vectors pAC1402, pAC1403 and pAC1404 containing ORFs of Clover_PUFa and Clover_PUFc, mRuby2_PUFa, respectively. .. These ORFs are then transferred to PB3-neo vector by recombining with pAC1119:PB3-neo(−)-pmaxDEST(+) by LR Clonase (Invitrogen) to create expression vectors pAC1360 (Clover_PUFa), pAC1381 (Clover_PUFc) and pAC1362 (mRuby2_PUFa).

    Article Title: Tissue‐like environments shape functional interactions of HIV ‐1 with immature dendritic cells
    Article Snippet: .. The plasmid pmRuby2.Vpr, encoding Vpr fused to mRuby2, was generated by excision of the eGFP coding region of peGFP.Vpr (McDonald et al , , a kind gift of Tom Hope) using AgeI and BsrGI restriction enzymes, followed by insertion of a PCR fragment encoding mRuby2 (Lam et al , ); amplified from pcDNA3‐mRuby2, a gift from Michael Lin (Addgene plasmid # 40260) and flanked by the same restriction sites. ..

    Cloning:

    Article Title: Three-component CRISPR/Cas complex system and uses thereof
    Article Snippet: A fusion PCR of 5′ fragment of NLSPUFa with 3′ fragment of NLSPUb was used to create pAC1358:pmax-NLSPUFc_VP64. p65HSF1 activator ORF was amplified from MS2-P65-HSF1_GFP (Addgene: 61423) with FseI PacI sites to replace VP64 fragment in pAC164 to create pAC1410:pmax-dCas9_p65HSF1, and replace VP64 in pAC1355 and pAC1358 to create pAC1393: pmax-NLSPUFa_p65HSF1 and pAC1411:pmax-NLSPUFc_p65HSF1, respectively. .. Clover and mRuby2 were amplified from pcDNA3-Clover (Addgene #40259) and pcDNA3-mRuby2 (Addgene #40260) respectively with primers containing SgrAI and FseI cloning site, ligated with various FseI-PUF-PacI amplified from the above pAC1356 ̃1358 and vector digested from pAC149:pCR8-dCas9VP160 (Addgene #48221) to create gateway donor vectors pAC1402, pAC1403 and pAC1404 containing ORFs of Clover_PUFa and Clover_PUFc, mRuby2_PUFa, respectively. .. These ORFs are then transferred to PB3-neo vector by recombining with pAC1119:PB3-neo(−)-pmaxDEST(+) by LR Clonase (Invitrogen) to create expression vectors pAC1360 (Clover_PUFa), pAC1381 (Clover_PUFc) and pAC1362 (mRuby2_PUFa).

    Expressing:

    Article Title: Exosomes as Conduits: Facilitating Hepatitis B Virus-Independent Hepatitis D Virus Transmission and Propagation in Hepatocytes
    Article Snippet: .. The plasmids pCIHD24 and pCIHD27 were used for the expression of S-HDAg and L-HDAg, respectively, as previously described [ , ]. pCIneo and peGFP were purchased from Promega and Clontech, respectively. pcDNA3 mRuby2 (named in pmRuby2) plasmid is from Addgene [ ]. .. Supernatants from transfected cells cultured in DMEM supplemented with 5% exosome-depleted serum were collected and clarified.

    other:

    Article Title: Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range
    Article Snippet: mRuby2 , pcDNA3-mRuby2 , 40260, Addgene, Cambridge, MA , 594.

    Generated:

    Article Title: Tissue‐like environments shape functional interactions of HIV ‐1 with immature dendritic cells
    Article Snippet: .. The plasmid pmRuby2.Vpr, encoding Vpr fused to mRuby2, was generated by excision of the eGFP coding region of peGFP.Vpr (McDonald et al , , a kind gift of Tom Hope) using AgeI and BsrGI restriction enzymes, followed by insertion of a PCR fragment encoding mRuby2 (Lam et al , ); amplified from pcDNA3‐mRuby2, a gift from Michael Lin (Addgene plasmid # 40260) and flanked by the same restriction sites. ..

    Polymerase Chain Reaction:

    Article Title: Tissue‐like environments shape functional interactions of HIV ‐1 with immature dendritic cells
    Article Snippet: .. The plasmid pmRuby2.Vpr, encoding Vpr fused to mRuby2, was generated by excision of the eGFP coding region of peGFP.Vpr (McDonald et al , , a kind gift of Tom Hope) using AgeI and BsrGI restriction enzymes, followed by insertion of a PCR fragment encoding mRuby2 (Lam et al , ); amplified from pcDNA3‐mRuby2, a gift from Michael Lin (Addgene plasmid # 40260) and flanked by the same restriction sites. ..



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    a Schematic overview of the experimental procedure: Day 1: preparation of thin (250 μm) and flat PAA gels; Day 2: surface functionalization with Sulfo-SANPAH and collagen coating in acidic solution; Day 3: seeding of cells transiently transfected with VinTS plasmid; Day 4: live cell imaging. b Imaging and data acquisition protocol. Step 1: stressed state bead and cell image acquisition (TFM imaging); Step 2: FLIM data acquisition of donor (Clover) channel (FRET imaging); Step3: intensity data of acceptor <t>(mRuby2)</t> channel; Step 4: relaxed state bead image acquired after the removal of the cells and the relaxation of the gel using a mild SDS solution. c Data processing workflow. Displacement fields generated from the overlay images of stressed and relaxed state bead images are calculated and subsequently converted into tractions. FRET efficiency is derived from the FRET trajectory between τ D and τ BG in the phasor space. FA structural and molecular properties are quantified from the acceptor intensity data after applying a binary FA mask. Illustrations were created using BioRender.com.
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    Addgene inc pcdna3 clover
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    Image Search Results


    a Schematic overview of the experimental procedure: Day 1: preparation of thin (250 μm) and flat PAA gels; Day 2: surface functionalization with Sulfo-SANPAH and collagen coating in acidic solution; Day 3: seeding of cells transiently transfected with VinTS plasmid; Day 4: live cell imaging. b Imaging and data acquisition protocol. Step 1: stressed state bead and cell image acquisition (TFM imaging); Step 2: FLIM data acquisition of donor (Clover) channel (FRET imaging); Step3: intensity data of acceptor (mRuby2) channel; Step 4: relaxed state bead image acquired after the removal of the cells and the relaxation of the gel using a mild SDS solution. c Data processing workflow. Displacement fields generated from the overlay images of stressed and relaxed state bead images are calculated and subsequently converted into tractions. FRET efficiency is derived from the FRET trajectory between τ D and τ BG in the phasor space. FA structural and molecular properties are quantified from the acceptor intensity data after applying a binary FA mask. Illustrations were created using BioRender.com.

    Journal: Communications Biology

    Article Title: Linking molecular tension and cellular tractions: a multiscale approach to focal adhesion mechanics

    doi: 10.1038/s42003-026-09514-0

    Figure Lengend Snippet: a Schematic overview of the experimental procedure: Day 1: preparation of thin (250 μm) and flat PAA gels; Day 2: surface functionalization with Sulfo-SANPAH and collagen coating in acidic solution; Day 3: seeding of cells transiently transfected with VinTS plasmid; Day 4: live cell imaging. b Imaging and data acquisition protocol. Step 1: stressed state bead and cell image acquisition (TFM imaging); Step 2: FLIM data acquisition of donor (Clover) channel (FRET imaging); Step3: intensity data of acceptor (mRuby2) channel; Step 4: relaxed state bead image acquired after the removal of the cells and the relaxation of the gel using a mild SDS solution. c Data processing workflow. Displacement fields generated from the overlay images of stressed and relaxed state bead images are calculated and subsequently converted into tractions. FRET efficiency is derived from the FRET trajectory between τ D and τ BG in the phasor space. FA structural and molecular properties are quantified from the acceptor intensity data after applying a binary FA mask. Illustrations were created using BioRender.com.

    Article Snippet: Plasmid DNA expressing free Clover (Addgene #40259) and mRuby2 (Addgene #40260) were gifts from Michael Lin .

    Techniques: Transfection, Plasmid Preparation, Live Cell Imaging, Imaging, Generated, Derivative Assay

    a Traction and FRET overlay images of representative cells on soft (4.5 kPa) and stiff (13 kPa) PAA substrate. Traction arrows are scaled with traction magnitude. b Comparison of cell surface area between soft and stiff PAA substrate. n = 93 and 85 cells. c Comparison of cell-averaged tractions between the soft and stiff substrate ( n = 51 and 38 cells, respectively). Each data point represents a single cell. d Illustration of the structure and working principle of Tension Sensing Module (TSMod) during donor excitation; Clover (C) and mRuby2 (R) FRET pair is separated with a synthetic flexible polypeptide (GGSGGS) 7 ; FRET efficiency remains high due to the module’s inability to support tension. e Illustration of the structure and working principle of Vinculin Tension Sensor (VinTS) used; TSMod is inserted between vinculin head domain (VinHD) and vinculin tail domain (VinTD); FRET efficiency decreases as a result of applied tension. f Averaged focal adhesion FRET efficiencies per cell for VinTS and force-insensitive control TSMod, compared on three different substrates ( n = 20, 93, 29, 85, 22, and 26 cells). Each data point represents a single cell. The middle line of violin plots represents the median, and the upper and lower lines indicate the 75th and 25th percentiles. Statistical analysis between two groups were performed using non-parametric Mann–Whitney U test (Wilcoxon rank-sum test) for ( b , c ), and unpaired two-tailed parametric Student’s t test for ( f ): * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Illustrations ( d , e ) were created using BioRender.com.

    Journal: Communications Biology

    Article Title: Linking molecular tension and cellular tractions: a multiscale approach to focal adhesion mechanics

    doi: 10.1038/s42003-026-09514-0

    Figure Lengend Snippet: a Traction and FRET overlay images of representative cells on soft (4.5 kPa) and stiff (13 kPa) PAA substrate. Traction arrows are scaled with traction magnitude. b Comparison of cell surface area between soft and stiff PAA substrate. n = 93 and 85 cells. c Comparison of cell-averaged tractions between the soft and stiff substrate ( n = 51 and 38 cells, respectively). Each data point represents a single cell. d Illustration of the structure and working principle of Tension Sensing Module (TSMod) during donor excitation; Clover (C) and mRuby2 (R) FRET pair is separated with a synthetic flexible polypeptide (GGSGGS) 7 ; FRET efficiency remains high due to the module’s inability to support tension. e Illustration of the structure and working principle of Vinculin Tension Sensor (VinTS) used; TSMod is inserted between vinculin head domain (VinHD) and vinculin tail domain (VinTD); FRET efficiency decreases as a result of applied tension. f Averaged focal adhesion FRET efficiencies per cell for VinTS and force-insensitive control TSMod, compared on three different substrates ( n = 20, 93, 29, 85, 22, and 26 cells). Each data point represents a single cell. The middle line of violin plots represents the median, and the upper and lower lines indicate the 75th and 25th percentiles. Statistical analysis between two groups were performed using non-parametric Mann–Whitney U test (Wilcoxon rank-sum test) for ( b , c ), and unpaired two-tailed parametric Student’s t test for ( f ): * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Illustrations ( d , e ) were created using BioRender.com.

    Article Snippet: Plasmid DNA expressing free Clover (Addgene #40259) and mRuby2 (Addgene #40260) were gifts from Michael Lin .

    Techniques: Comparison, Single Cell, Control, MANN-WHITNEY, Two Tailed Test