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Addgene inc second generation car targeting cd19

Top-performing LNPs generate <t>CAR-Jurkat</t> cells with high efficiency. Jurkat cells were transfected with top-performing LNPs using 420 ng/60,000 cells, collected after 3 days and co-cultured at 1:1 or 1:0 ratios with Raji cells. ( A ) Schematic of Jurkat cell transfection with LNP-CAR and effector cell activation after co-culture with Raji cells. ( B ) Representative histogram of Jurkat and Raji cells distinguished based on the expression of <t>CD19.</t> ( C ) Expression of activation markers PD-1 and CD69 in Jurkat cells expressing <t>CAR-CD19BBz.</t> ( D and E ) Activation of Jurkat cells expressing CAR-CD19BBz measured by mean fluorescence intensity (MFI) of CD69 ( D ) and PD-1 ( E ) after 24 hours co-cultured with Raji cells. * p<0.05; ** p<0.01; ***p<0.001; **** p<0.0001 Not significant (ns) p>0.05 versus LNP-9-GN1; “a” = p<0.001 compared to 1:1; “b” = p<0.0001 compared to 1:1; evaluated by one-way ANOVA with n=3. ( F and G ) Activation of Jurkat cells transfected with the optimized formulation LNP-9 24 hours after co-culture with Raji cells at different effector-to-target ratios. Activation was assessed by the expression of CD69 ( F ) and PD-1 ( G ). *p<0.05; **p<0.01; ***p<0.001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=3. Graphs represent mean ± SD.

Second Generation Car Targeting Cd19, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 27 article reviews

second generation car targeting cd19 - by Bioz Stars, 2026-04

93/100 stars

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1) Product Images from "Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells"

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S424723

Top-performing LNPs generate CAR-Jurkat cells with high efficiency. Jurkat cells were transfected with top-performing LNPs using 420 ng/60,000 cells, collected after 3 days and co-cultured at 1:1 or 1:0 ratios with Raji cells. ( A ) Schematic of Jurkat cell transfection with LNP-CAR and effector cell activation after co-culture with Raji cells. ( B ) Representative histogram of Jurkat and Raji cells distinguished based on the expression of CD19. ( C ) Expression of activation markers PD-1 and CD69 in Jurkat cells expressing CAR-CD19BBz. ( D and E ) Activation of Jurkat cells expressing CAR-CD19BBz measured by mean fluorescence intensity (MFI) of CD69 ( D ) and PD-1 ( E ) after 24 hours co-cultured with Raji cells. * p<0.05; ** p<0.01; ***p<0.001; **** p<0.0001 Not significant (ns) p>0.05 versus LNP-9-GN1; “a” = p<0.001 compared to 1:1; “b” = p<0.0001 compared to 1:1; evaluated by one-way ANOVA with n=3. ( F and G ) Activation of Jurkat cells transfected with the optimized formulation LNP-9 24 hours after co-culture with Raji cells at different effector-to-target ratios. Activation was assessed by the expression of CD69 ( F ) and PD-1 ( G ). *p<0.05; **p<0.01; ***p<0.001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=3. Graphs represent mean ± SD.

Figure Legend Snippet: Top-performing LNPs generate CAR-Jurkat cells with high efficiency. Jurkat cells were transfected with top-performing LNPs using 420 ng/60,000 cells, collected after 3 days and co-cultured at 1:1 or 1:0 ratios with Raji cells. ( A ) Schematic of Jurkat cell transfection with LNP-CAR and effector cell activation after co-culture with Raji cells. ( B ) Representative histogram of Jurkat and Raji cells distinguished based on the expression of CD19. ( C ) Expression of activation markers PD-1 and CD69 in Jurkat cells expressing CAR-CD19BBz. ( D and E ) Activation of Jurkat cells expressing CAR-CD19BBz measured by mean fluorescence intensity (MFI) of CD69 ( D ) and PD-1 ( E ) after 24 hours co-cultured with Raji cells. * p<0.05; ** p<0.01; ***p<0.001; **** p<0.0001 Not significant (ns) p>0.05 versus LNP-9-GN1; “a” = p<0.001 compared to 1:1; “b” = p<0.0001 compared to 1:1; evaluated by one-way ANOVA with n=3. ( F and G ) Activation of Jurkat cells transfected with the optimized formulation LNP-9 24 hours after co-culture with Raji cells at different effector-to-target ratios. Activation was assessed by the expression of CD69 ( F ) and PD-1 ( G ). *p<0.05; **p<0.01; ***p<0.001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=3. Graphs represent mean ± SD.

Techniques Used: Transfection, Cell Culture, Activation Assay, Co-Culture Assay, Expressing, Fluorescence, Formulation

LNP-9-CAR promotes functional delivery of pDNA to primary human T cells. ( A ) Schematic of primary human T cell transfection with LNP-9-CAR ( B ) Representative plots of CAR-CD19BBz expression 3 days after transfection of primary human T cells with LNP-9-CAR. ( C ) Cell viability of CD3 + cells 3 days after transfection with LNP-9-CAR. Not significant (ns) p>0.05 evaluated by unpaired t -test compared to NTC. ( D ) Frequency of Live/CD3 + /CAR-CD19BBz + T cells analyzed 3 days after transfection with LNP-9-CAR. ***p<0.001 evaluated by unpaired t -test compared to NTC. ( E ) Representative plots of T cell subsets expressing CAR-CD19BBz 3 days after transfection with LNP-9-CAR identified by the expression of CD4 and CD8. ( F ) Frequency of Live/CD3 + /CAR-CD19BBz + CD4 + and CD8 + T cells analyzed 3 days after transfection with LNP-9-CAR. **p<0.01 evaluated by unpaired t -test compared to NTC. ( G ) Schematic of T cell transfection with LNP-9-CAR and tumor cell specific killing in co-culture with Raji cells. ( H ) Representative plots of Annexin V/7AAD labeled cells gated on the CD3 − /CD19 + population. Dead cells are labeled as double positive for both markers. ( I ) Results of specific killing of CD19 + 24 hours after coplating with CAR T cells or non-transfected cells at different ratios. *p<0.05; ****p<0.0001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=4 donors. NTC = non-transfected control. Graphs represent mean ± SD.

Figure Legend Snippet: LNP-9-CAR promotes functional delivery of pDNA to primary human T cells. ( A ) Schematic of primary human T cell transfection with LNP-9-CAR ( B ) Representative plots of CAR-CD19BBz expression 3 days after transfection of primary human T cells with LNP-9-CAR. ( C ) Cell viability of CD3 + cells 3 days after transfection with LNP-9-CAR. Not significant (ns) p>0.05 evaluated by unpaired t -test compared to NTC. ( D ) Frequency of Live/CD3 + /CAR-CD19BBz + T cells analyzed 3 days after transfection with LNP-9-CAR. ***p<0.001 evaluated by unpaired t -test compared to NTC. ( E ) Representative plots of T cell subsets expressing CAR-CD19BBz 3 days after transfection with LNP-9-CAR identified by the expression of CD4 and CD8. ( F ) Frequency of Live/CD3 + /CAR-CD19BBz + CD4 + and CD8 + T cells analyzed 3 days after transfection with LNP-9-CAR. **p<0.01 evaluated by unpaired t -test compared to NTC. ( G ) Schematic of T cell transfection with LNP-9-CAR and tumor cell specific killing in co-culture with Raji cells. ( H ) Representative plots of Annexin V/7AAD labeled cells gated on the CD3 − /CD19 + population. Dead cells are labeled as double positive for both markers. ( I ) Results of specific killing of CD19 + 24 hours after coplating with CAR T cells or non-transfected cells at different ratios. *p<0.05; ****p<0.0001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=4 donors. NTC = non-transfected control. Graphs represent mean ± SD.

Techniques Used: Functional Assay, Transfection, Expressing, Co-Culture Assay, Labeling, Control

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Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: Top-performing LNPs generate CAR-Jurkat cells with high efficiency. Jurkat cells were transfected with top-performing LNPs using 420 ng/60,000 cells, collected after 3 days and co-cultured at 1:1 or 1:0 ratios with Raji cells. ( A ) Schematic of Jurkat cell transfection with LNP-CAR and effector cell activation after co-culture with Raji cells. ( B ) Representative histogram of Jurkat and Raji cells distinguished based on the expression of CD19. ( C ) Expression of activation markers PD-1 and CD69 in Jurkat cells expressing CAR-CD19BBz. ( D and E ) Activation of Jurkat cells expressing CAR-CD19BBz measured by mean fluorescence intensity (MFI) of CD69 ( D ) and PD-1 ( E ) after 24 hours co-cultured with Raji cells. * p<0.05; ** p<0.01; ***p<0.001; **** p<0.0001 Not significant (ns) p>0.05 versus LNP-9-GN1; “a” = p<0.001 compared to 1:1; “b” = p<0.0001 compared to 1:1; evaluated by one-way ANOVA with n=3. ( F and G ) Activation of Jurkat cells transfected with the optimized formulation LNP-9 24 hours after co-culture with Raji cells at different effector-to-target ratios. Activation was assessed by the expression of CD69 ( F ) and PD-1 ( G ). *p<0.05; **p<0.01; ***p<0.001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=3. Graphs represent mean ± SD.

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Transfection, Cell Culture, Activation Assay, Co-Culture Assay, Expressing, Fluorescence, Formulation

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: LNP-9-CAR promotes functional delivery of pDNA to primary human T cells. ( A ) Schematic of primary human T cell transfection with LNP-9-CAR ( B ) Representative plots of CAR-CD19BBz expression 3 days after transfection of primary human T cells with LNP-9-CAR. ( C ) Cell viability of CD3 + cells 3 days after transfection with LNP-9-CAR. Not significant (ns) p>0.05 evaluated by unpaired t -test compared to NTC. ( D ) Frequency of Live/CD3 + /CAR-CD19BBz + T cells analyzed 3 days after transfection with LNP-9-CAR. ***p<0.001 evaluated by unpaired t -test compared to NTC. ( E ) Representative plots of T cell subsets expressing CAR-CD19BBz 3 days after transfection with LNP-9-CAR identified by the expression of CD4 and CD8. ( F ) Frequency of Live/CD3 + /CAR-CD19BBz + CD4 + and CD8 + T cells analyzed 3 days after transfection with LNP-9-CAR. **p<0.01 evaluated by unpaired t -test compared to NTC. ( G ) Schematic of T cell transfection with LNP-9-CAR and tumor cell specific killing in co-culture with Raji cells. ( H ) Representative plots of Annexin V/7AAD labeled cells gated on the CD3 − /CD19 + population. Dead cells are labeled as double positive for both markers. ( I ) Results of specific killing of CD19 + 24 hours after coplating with CAR T cells or non-transfected cells at different ratios. *p<0.05; ****p<0.0001; Not significant (ns) p>0.05 evaluated by two-way ANOVA with n=4 donors. NTC = non-transfected control. Graphs represent mean ± SD.

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Functional Assay, Transfection, Expressing, Co-Culture Assay, Labeling, Control

Composition and Characterization of pDNA-ZsGreen Encapsulated in LNPs with Different DOPE and Cholesterol Ratios

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: Composition and Characterization of pDNA-ZsGreen Encapsulated in LNPs with Different DOPE and Cholesterol Ratios

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques:

Composition and Characterization of pDNA-CAR-CD19BBz Encapsulated LNPs with Different DOPE and Cholesterol Ratios

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: Composition and Characterization of pDNA-CAR-CD19BBz Encapsulated LNPs with Different DOPE and Cholesterol Ratios

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Encapsulation

DNA loaded LNPs induce transient expression of CAR-CD19BBz in Jurkat cells. ( A ) Schematic of LNP-CAR mediated transfection of Jurkat cells. ( B ) CAR-CD19BBz expression in live (PI − ) Jurkat cells 3 days after transfection with LNP-5-CAR with different amounts of pDNA identifying the best concentration to be used. * p<0.05; **** p<0.0001 versus 420 ng/ 60,000 cells evaluated by one-way ANOVA with n=3. ( C ) Cell viability of Jurkat cells assessed by the frequency of PI − cells after transfection with LNP-5-CAR. ( D ) CAR-CD19BBz expression over time in Jurkat cells treated with 420 ng/60,000 cells of LNP-5-CAR confirming the transient expression of CAR in these cells. ** p<0.01; ***p<0.001; ****p<0.0001 versus NTC on the same timepoint evaluated by one-way ANOVA with n=3. ( E ) Cell viability of Jurkat cells over time. * p<0.05; ****p<0.0001; Not significant (ns) p>0.05 versus NTC on the same timepoint evaluated by one-way ANOVA with n=3.

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: DNA loaded LNPs induce transient expression of CAR-CD19BBz in Jurkat cells. ( A ) Schematic of LNP-CAR mediated transfection of Jurkat cells. ( B ) CAR-CD19BBz expression in live (PI − ) Jurkat cells 3 days after transfection with LNP-5-CAR with different amounts of pDNA identifying the best concentration to be used. * p<0.05; **** p<0.0001 versus 420 ng/ 60,000 cells evaluated by one-way ANOVA with n=3. ( C ) Cell viability of Jurkat cells assessed by the frequency of PI − cells after transfection with LNP-5-CAR. ( D ) CAR-CD19BBz expression over time in Jurkat cells treated with 420 ng/60,000 cells of LNP-5-CAR confirming the transient expression of CAR in these cells. ** p<0.01; ***p<0.001; ****p<0.0001 versus NTC on the same timepoint evaluated by one-way ANOVA with n=3. ( E ) Cell viability of Jurkat cells over time. * p<0.05; ****p<0.0001; Not significant (ns) p>0.05 versus NTC on the same timepoint evaluated by one-way ANOVA with n=3.

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Expressing, Transfection, Concentration Assay

Higher DOPE molar ratios facilitate efficient transfection with CAR-CD19BBz in Jurkat cells. ( A ) Schematic of the components used to prepare LNPs by microfluidic mixing. ( B ) Schematic of LNP-CAR screening using Jurkat cells. ( C ) Hydrodynamic diameter (Size) and polydispersity index (PDI) of different LNPs encapsulating CAR-CD19BBz pDNA. ( D ) Zeta potential of different LNPs. ( E ) Representative cryo-TEM of LNPs encapsulating pDNA. ( F ) CAR-CD19BBz expression in live (PI − ) Jurkat cells 3 days after transfection with different LNPs identifying top-performing LNPs. **** p<0.0001 versus NTC evaluated by one-way ANOVA with n=3. ( G ) Cell viability of Jurkat cells assessed by the frequency of PI − cells after transfection with top-performing LNPs **** p<0.0001 versus NTC; evaluated by one-way ANOVA with n=3. ( H ) Representative histograms of CAR-CB19BBz expression after transfection. NTC = non-transfected control; EP = Electroporation; Lipo = Lipofectamine. Graphs represent mean ± SD. ( I ) CAR-CD19BBz expression measured by mean fluorescence intensity (MFI) after transfection with top-performing LNPs and controls. **p<0.01; ****p<0.0001; Not significant (ns) p>0.05 versus NTC evaluated by one-way ANOVA with n=3. “a” = p<0.001 compared to EP; “b” = p<0.001 compared to Lipo evaluated by one-way ANOVA with n=3. NTC = non-transfected control; Graphs represent mean ± SD.

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Figure Lengend Snippet: Higher DOPE molar ratios facilitate efficient transfection with CAR-CD19BBz in Jurkat cells. ( A ) Schematic of the components used to prepare LNPs by microfluidic mixing. ( B ) Schematic of LNP-CAR screening using Jurkat cells. ( C ) Hydrodynamic diameter (Size) and polydispersity index (PDI) of different LNPs encapsulating CAR-CD19BBz pDNA. ( D ) Zeta potential of different LNPs. ( E ) Representative cryo-TEM of LNPs encapsulating pDNA. ( F ) CAR-CD19BBz expression in live (PI − ) Jurkat cells 3 days after transfection with different LNPs identifying top-performing LNPs. **** p<0.0001 versus NTC evaluated by one-way ANOVA with n=3. ( G ) Cell viability of Jurkat cells assessed by the frequency of PI − cells after transfection with top-performing LNPs **** p<0.0001 versus NTC; evaluated by one-way ANOVA with n=3. ( H ) Representative histograms of CAR-CB19BBz expression after transfection. NTC = non-transfected control; EP = Electroporation; Lipo = Lipofectamine. Graphs represent mean ± SD. ( I ) CAR-CD19BBz expression measured by mean fluorescence intensity (MFI) after transfection with top-performing LNPs and controls. **p<0.01; ****p<0.0001; Not significant (ns) p>0.05 versus NTC evaluated by one-way ANOVA with n=3. “a” = p<0.001 compared to EP; “b” = p<0.001 compared to Lipo evaluated by one-way ANOVA with n=3. NTC = non-transfected control; Graphs represent mean ± SD.

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Transfection, Zeta Potential Analyzer, Expressing, Control, Electroporation, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells

doi: 10.2147/IJN.S424723

Article Snippet: We encapsulated a pDNA encoding for a second-generation CAR targeting CD19 (CAR-CD19BBz, Addgene no. 135992) in our lead formulation LNP-5 (LNP-5-CAR).

Techniques: Functional Assay, Transfection, Expressing, Co-Culture Assay, Labeling, Control

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