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ms2 bacteriophage coat protein tandem dimer  (Addgene inc)


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    Addgene inc ms2 bacteriophage coat protein tandem dimer
    Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
    Ms2 Bacteriophage Coat Protein Tandem Dimer, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ms2 bacteriophage coat protein tandem dimer/product/Addgene inc
    Average 95 stars, based on 153 article reviews
    ms2 bacteriophage coat protein tandem dimer - by Bioz Stars, 2026-02
    95/100 stars

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    1) Product Images from "A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery"

    Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

    Journal: Journal of Extracellular Vesicles

    doi: 10.3402/jev.v5.31027

    Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
    Figure Legend Snippet: Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.

    Techniques Used: RNA Binding Assay, Derivative Assay, Expressing, Reverse Transcription, Amplification, Transfection, Incubation, Standard Deviation

    Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.
    Figure Legend Snippet: Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.

    Techniques Used: Quantitative Proteomics, Western Blot, Standard Deviation

    Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.
    Figure Legend Snippet: Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.

    Techniques Used: Standard Deviation, Fluorescence, Comparison, Transfection



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    ms2 bacteriophage coat protein tandem dimer  (Addgene inc)
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    Addgene inc ms2 bacteriophage coat protein tandem dimer
    Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA <t>+/−MS2”</t> is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.
    Ms2 Bacteriophage Coat Protein Tandem Dimer, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ms2 bacteriophage coat protein tandem dimer/product/Addgene inc
    Average 95 stars, based on 1 article reviews
    ms2 bacteriophage coat protein tandem dimer - by Bioz Stars, 2026-02
    95/100 stars
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    Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.

    Journal: Journal of Extracellular Vesicles

    Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

    doi: 10.3402/jev.v5.31027

    Figure Lengend Snippet: Evaluation of RNA loading into EVs via the TAMEL platform. (a) This cartoon summarizes the concept of facilitating active loading of cargo RNA into EVs via our TAMEL platform. A TAMEL EV-loading protein comprises an EV-enriched protein (EEP, blue) fused to an RNA-binding domain (RBD, green), which localizes to EVs. Actively loaded RNA (green) contains a motif that binds to the RBD, resulting in enhanced loading into EVs relative to passively loaded RNA (orange). (b) RNA cargo design impacts active loading. The “fold enrichment of cargo mRNA +/−MS2” is defined as the ratio of cargo RNA/GAPDH mRNA in EVs derived from cells expressing Lamp2b–MS2–HA divided by the same RNA ratio in EVs derived from cells expressing Lamp2b–HA. All experiments were performed in biological triplicates. (c) Cartoon illustrating the 3′ RNA fragment analysis technique. Cargo RNA is first reverse transcribed using an oligo dT primer, and amplicons corresponding to the RNA 5′ or 3′ ends (the latter is located ~500 bases upstream of the polyA site) are then quantified by qPCR using the primer pairs indicated. Note that the amplicon near the RNA 3′ end will be present in cDNA derived from both full-length RNA and 3′ RNA fragments. (d) Analysis of 3′ RNA fragment loading into EVs. Cargo RNA levels were quantified as depicted in panel c and normalized to GAPDH. Passive loading: cells transfected with Lamp2b–HA; active loading: cells transfected with Lamp2b–MS2–HA. (e) Full-length RNA and 3′ fragment RNA levels in EVs were quantified following incubation at 37°C; experiments were performed in technical duplicate with a biological replicate shown in Supplementary Fig. 2a. Error bars indicate 1 standard deviation, throughout. MVB, multivesicular body.

    Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

    Techniques: RNA Binding Assay, Derivative Assay, Expressing, Reverse Transcription, Amplification, Transfection, Incubation, Standard Deviation

    Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.

    Journal: Journal of Extracellular Vesicles

    Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

    doi: 10.3402/jev.v5.31027

    Figure Lengend Snippet: Impact of EEP choice on TAMEL-mediated active RNA loading into vesicles. (a) Effects of EEP choice on cargo RNA loading into EVs. Experiments were performed in biological triplicate. (b) Effects of EEP choice on cargo loading into gesicles. Experiments were performed in biological triplicate. (c) Protein abundance was quantified by densitometry analysis of anti-HA western blots (Supplementary Fig. 3), and each blot was internally normalized by the intensity for VSVG–MS2–HA in gesicles (maximal intensity case). This experiment was performed in biological duplicate. The y-axis is in log scale to enable visualization of all values. Error bars indicate 1 standard deviation, throughout.

    Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

    Techniques: Quantitative Proteomics, Western Blot, Standard Deviation

    Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.

    Journal: Journal of Extracellular Vesicles

    Article Title: A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery

    doi: 10.3402/jev.v5.31027

    Figure Lengend Snippet: Comparative analysis of dTomato delivery by actively or passively loaded vesicles. For panels a–c, the cartoons at left summarize the experimental designs, and in the panels at right, each data point represents the average of duplicate wells of cells treated with the same type of vesicle. Error bars indicate 1 standard deviation. “Normalized fluorescence” is defined as the mean fluorescence of cells receiving vesicles divided by the mean fluorescence of cells receiving a medium change only. (a) Time course of EV delivery to cells. Grey squares: CD63–HA EVs; orange circles: CD63–MS2–HA EVs. The solid arrow represents cells that received a medium change after 4 h of EV treatment. The dashed arrow indicates that cells did not receive a medium change. Statistically significant differences (p<0.05, not shown for clarity): CD63–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3a. (b) Time course of gesicle delivery to cells. Purple squares: VSVG–HA gesicles; green circles: VSVG–MS2–HA gesicles. Solid and dashed arrows carry the same meaning as in panel (a). Statistically significant differences (p < 0.05, not shown for clarity): VSVG–HA versus VSVG–MS2–HA at 4 and 16 h (comparisons were made for each time point), VSVG–HA +/− medium change, and VSVG–MS2–HA +/− medium change. An independent repeat of this experiment is shown in Supplementary Fig. 3c. (c) Comparison of delivery by gesicles from cells transfected with VSVG–HA (purple), VSVG–MS2–HA (green) or a 50:50 mix of VSVG–HA and VSVG–MS2–HA (hybrid gesicles, magenta). (d) dTomato RNA levels (normalized to GAPDH) in VSVG–HA gesicles (purple), VSVG–MS2–HA gesicles (green) or hybrid gesicles (magenta). Error bars indicate 1 standard deviation of technical duplicate samples. *Significant difference was evaluated with a Student's t -test using a cut-off of p < 0.05.

    Article Snippet: The pMS2-GFP plasmid, which encodes the MS2 bacteriophage coat protein tandem dimer (mutant d1FG and V29I ( , )), was obtained from Addgene (plasmid #27121), deposited by Robert Singer ( ).

    Techniques: Standard Deviation, Fluorescence, Comparison, Transfection