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expression plasmid pmscv puro ires gfp 64  (Addgene inc)


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    Addgene inc expression plasmid pmscv puro ires gfp 64
    Expression Plasmid Pmscv Puro Ires Gfp 64, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pmscv-vector/pMSCV+PIG+(Puro+IRES+GFP+empty+vector)+(Plasmid+%2321654)/10__1016_slash_j__isci__2026__115182-641-41-45
    Average 93 stars, based on 77 article reviews
    expression plasmid pmscv puro ires gfp 64 - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Construct:

    Article Title: Mitochondria-derived PEP licenses glycerolipid synthesis via SLC25A35
    Article Snippet: .. Mouse Slc25a35 cDNA constructs were amplified from mouse inguinal WAT and inserted into a blasticidin-resistant pMSCV vector (75085, Addgene). ..

    Article Title: A Stress-Induced PI3P Complex Controls Cell Autophagy in Erythroid Precursors
    Article Snippet: .. Plasmid Constructs 61 HA-tagged full-length Samd14, Samd14 lacking the SAM domain (S14ΔSAM), Samd14 lacking the capping 62 protein binding (CPB) domain (Samd14 ΔCPB), and Samd14 lacking both SAM and CPB domains 63 (S14ΔCS) (Ray et al., 2022) were cloned into mammalian expression plasmid pMSCV-Puro-IRES-GFP 64 (Addgene #21654) using BglII and EcoRI restriction digestion sites. ..

    Amplification:

    Article Title: Mitochondria-derived PEP licenses glycerolipid synthesis via SLC25A35
    Article Snippet: .. Mouse Slc25a35 cDNA constructs were amplified from mouse inguinal WAT and inserted into a blasticidin-resistant pMSCV vector (75085, Addgene). ..

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Plasmid Preparation:

    Article Title: Mitochondria-derived PEP licenses glycerolipid synthesis via SLC25A35
    Article Snippet: .. Mouse Slc25a35 cDNA constructs were amplified from mouse inguinal WAT and inserted into a blasticidin-resistant pMSCV vector (75085, Addgene). ..

    Article Title: Syk activation during FcγR-mediated phagocytosis involves Syk palmitoylation and desulfenylation.
    Article Snippet: .. The mSyk expression vector was obtained from Addgene (#50045). .. FLAG-mSyk was prepared by inserting a FLAG epitope (DYKDDDDK) upstream of Syk coding sequence in the pCi vector (Promega).

    Article Title: Syk activation during FcγR-mediated phagocytosis involves Syk palmitoylation and desulfenylation
    Article Snippet: .. The mSyk expression vector was obtained from Addgene (#50045). .. FLAG-mSyk was prepared by inserting a FLAG epitope (DYKDDDDK) upstream of Syk coding sequence in the pCi vector (Promega).

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: A Stress-Induced PI3P Complex Controls Cell Autophagy in Erythroid Precursors
    Article Snippet: .. Plasmid Constructs 61 HA-tagged full-length Samd14, Samd14 lacking the SAM domain (S14ΔSAM), Samd14 lacking the capping 62 protein binding (CPB) domain (Samd14 ΔCPB), and Samd14 lacking both SAM and CPB domains 63 (S14ΔCS) (Ray et al., 2022) were cloned into mammalian expression plasmid pMSCV-Puro-IRES-GFP 64 (Addgene #21654) using BglII and EcoRI restriction digestion sites. ..

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Mutagenesis:

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators
    Article Snippet: All cell lines were Mycoplasma-free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-701; MycoAlert Assay Control Set, Lonza, LT07-518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV-IRES-YFP II (RRID:Addgene_52108) to generate MSCV-YFP-hIDH2-R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL-243) were transduced with the MSCV-YFP-hIDH2-R140Q vector, and mutant YFP + cells were subject to fluorescence-activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators.
    Article Snippet: All cell lines were Mycoplasma- free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07- 701; MycoAlert Assay Control Set, Lonza, LT07- 518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV- IRESYFP II (RRID:Addgene_52108) to generate MSCV- YFP- hIDH2R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL- 243) were transduced with the MSCV- YFP- hIDH2- R140Q vector, and mutant YFP+ cells were subject to fluorescence- activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_ 016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Clone Assay:

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators
    Article Snippet: All cell lines were Mycoplasma-free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-701; MycoAlert Assay Control Set, Lonza, LT07-518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV-IRES-YFP II (RRID:Addgene_52108) to generate MSCV-YFP-hIDH2-R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL-243) were transduced with the MSCV-YFP-hIDH2-R140Q vector, and mutant YFP + cells were subject to fluorescence-activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators.
    Article Snippet: All cell lines were Mycoplasma- free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07- 701; MycoAlert Assay Control Set, Lonza, LT07- 518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV- IRESYFP II (RRID:Addgene_52108) to generate MSCV- YFP- hIDH2R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL- 243) were transduced with the MSCV- YFP- hIDH2- R140Q vector, and mutant YFP+ cells were subject to fluorescence- activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_ 016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: A Stress-Induced PI3P Complex Controls Cell Autophagy in Erythroid Precursors
    Article Snippet: .. Plasmid Constructs 61 HA-tagged full-length Samd14, Samd14 lacking the SAM domain (S14ΔSAM), Samd14 lacking the capping 62 protein binding (CPB) domain (Samd14 ΔCPB), and Samd14 lacking both SAM and CPB domains 63 (S14ΔCS) (Ray et al., 2022) were cloned into mammalian expression plasmid pMSCV-Puro-IRES-GFP 64 (Addgene #21654) using BglII and EcoRI restriction digestion sites. ..

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Expressing:

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators
    Article Snippet: All cell lines were Mycoplasma-free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-701; MycoAlert Assay Control Set, Lonza, LT07-518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV-IRES-YFP II (RRID:Addgene_52108) to generate MSCV-YFP-hIDH2-R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL-243) were transduced with the MSCV-YFP-hIDH2-R140Q vector, and mutant YFP + cells were subject to fluorescence-activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators.
    Article Snippet: All cell lines were Mycoplasma- free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07- 701; MycoAlert Assay Control Set, Lonza, LT07- 518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV- IRESYFP II (RRID:Addgene_52108) to generate MSCV- YFP- hIDH2R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL- 243) were transduced with the MSCV- YFP- hIDH2- R140Q vector, and mutant YFP+ cells were subject to fluorescence- activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_ 016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Syk activation during FcγR-mediated phagocytosis involves Syk palmitoylation and desulfenylation.
    Article Snippet: .. The mSyk expression vector was obtained from Addgene (#50045). .. FLAG-mSyk was prepared by inserting a FLAG epitope (DYKDDDDK) upstream of Syk coding sequence in the pCi vector (Promega).

    Article Title: Syk activation during FcγR-mediated phagocytosis involves Syk palmitoylation and desulfenylation
    Article Snippet: .. The mSyk expression vector was obtained from Addgene (#50045). .. FLAG-mSyk was prepared by inserting a FLAG epitope (DYKDDDDK) upstream of Syk coding sequence in the pCi vector (Promega).

    Article Title: A Stress-Induced PI3P Complex Controls Cell Autophagy in Erythroid Precursors
    Article Snippet: .. Plasmid Constructs 61 HA-tagged full-length Samd14, Samd14 lacking the SAM domain (S14ΔSAM), Samd14 lacking the capping 62 protein binding (CPB) domain (Samd14 ΔCPB), and Samd14 lacking both SAM and CPB domains 63 (S14ΔCS) (Ray et al., 2022) were cloned into mammalian expression plasmid pMSCV-Puro-IRES-GFP 64 (Addgene #21654) using BglII and EcoRI restriction digestion sites. ..

    Retroviral:

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators
    Article Snippet: All cell lines were Mycoplasma-free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-701; MycoAlert Assay Control Set, Lonza, LT07-518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV-IRES-YFP II (RRID:Addgene_52108) to generate MSCV-YFP-hIDH2-R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL-243) were transduced with the MSCV-YFP-hIDH2-R140Q vector, and mutant YFP + cells were subject to fluorescence-activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Synergistic intragenic epigenetic deregulation by IDH2 and SRSF2 mutations causes mis-splicing of key transcriptional regulators.
    Article Snippet: All cell lines were Mycoplasma- free and routinely tested by Antibody and Bioresource Core at the Memorial Sloan Kettering Cancer Center (MSKCC) (MycoAlert Mycoplasma Detection Kit, Lonza, LT07- 701; MycoAlert Assay Control Set, Lonza, LT07- 518). .. The human IDH2 gene with R140Q mutation was cloned into a yellow fluorescent protein (YFP) expressing vector pMSCV- IRESYFP II (RRID:Addgene_52108) to generate MSCV- YFP- hIDH2R140Q retroviral vector. .. For generation of the transgenic human IDH2 R140Q cell line, parental K562 cells from ATCC (#CCL- 243) were transduced with the MSCV- YFP- hIDH2- R140Q vector, and mutant YFP+ cells were subject to fluorescence- activated cell sorting (FACS) using a FACSAria III Cell Sorter (BD Biosciences, RRID:SCR_ 016695) to isolate the K562 IDH2 R140Q mutant expressing cell line.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Polymerase Chain Reaction:

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Ligation:

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model
    Article Snippet: .. The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP. .. A synthetic gene of human/murine hybrid EGFR (BioInnLabs, Rostov-on-Don, Russia) was inserted into the pLV3 lentiviral vector (Clontech, San Jose, CA, USA) with the same method.

    Protein Binding:

    Article Title: A Stress-Induced PI3P Complex Controls Cell Autophagy in Erythroid Precursors
    Article Snippet: .. Plasmid Constructs 61 HA-tagged full-length Samd14, Samd14 lacking the SAM domain (S14ΔSAM), Samd14 lacking the capping 62 protein binding (CPB) domain (Samd14 ΔCPB), and Samd14 lacking both SAM and CPB domains 63 (S14ΔCS) (Ray et al., 2022) were cloned into mammalian expression plasmid pMSCV-Puro-IRES-GFP 64 (Addgene #21654) using BglII and EcoRI restriction digestion sites. ..



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    Image Search Results


    Design of human/murine hybrid epidermal growth factor receptor (EGFR) and EGFR murine chimeric antigen receptor (CAR). ( A ) Schematic of the human/murine hybrid EGFR with cetuximab-binding substitutions (G442S, R467K, M491I, N492S, K497N) and its engagement by a cetuximab-derived single-chain variable fragment (scFv) on mCAR T cells. ( B ) The retroviral vector encoding the EGFR-specific murine CAR and green fluorescent protein (GFP); I—domain I; II—domain II; III—domain III; IV—domain IV.

    Journal: International Journal of Molecular Sciences

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model

    doi: 10.3390/ijms27010189

    Figure Lengend Snippet: Design of human/murine hybrid epidermal growth factor receptor (EGFR) and EGFR murine chimeric antigen receptor (CAR). ( A ) Schematic of the human/murine hybrid EGFR with cetuximab-binding substitutions (G442S, R467K, M491I, N492S, K497N) and its engagement by a cetuximab-derived single-chain variable fragment (scFv) on mCAR T cells. ( B ) The retroviral vector encoding the EGFR-specific murine CAR and green fluorescent protein (GFP); I—domain I; II—domain II; III—domain III; IV—domain IV.

    Article Snippet: The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP.

    Techniques: Binding Assay, Derivative Assay, Retroviral, Plasmid Preparation

    Generation of murine EGFR CAR T cells. ( A ) Workflow for EGFR mCAR T cell production, including T cell isolation, activation, retroviral transduction, and subsequent in vitro and in vivo experiments created with BioRender ( https://www.biorender.com/ (accessed on 1 September 2025)). ( B ) Representative flow cytometry histogram showing the transduction efficiency of murine T cells with the EGFR-targeted murine CAR construct. ( C ) Summary of independent transduction preparations: percentage of GFP + cells in each preparation. ( D ) Expansion ( left ) and viability ( right ) of EGFR mCAR T cells compared to non-transduced T cells over 14 days of culture. * p < 0.05 and *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: Pre-Treatment with Dacarbazine Sensitizes B16 Melanoma to CAR T Cell Therapy in Syngeneic Mouse Model

    doi: 10.3390/ijms27010189

    Figure Lengend Snippet: Generation of murine EGFR CAR T cells. ( A ) Workflow for EGFR mCAR T cell production, including T cell isolation, activation, retroviral transduction, and subsequent in vitro and in vivo experiments created with BioRender ( https://www.biorender.com/ (accessed on 1 September 2025)). ( B ) Representative flow cytometry histogram showing the transduction efficiency of murine T cells with the EGFR-targeted murine CAR construct. ( C ) Summary of independent transduction preparations: percentage of GFP + cells in each preparation. ( D ) Expansion ( left ) and viability ( right ) of EGFR mCAR T cells compared to non-transduced T cells over 14 days of culture. * p < 0.05 and *** p < 0.001.

    Article Snippet: The synthetic CAR gene was cloned by PCR amplification with primers containing restriction sites, restriction digest, and ligation (restriction enzymes and ligase from New England Biolabs, Ipswich, MA, USA) into the retroviral vector pMSCV (a gift from Dr. Reya, Addgene, #20672) right before IRES followed by the reporter gene of GFP.

    Techniques: Cell Isolation, Activation Assay, Retroviral, Transduction, In Vitro, In Vivo, Flow Cytometry, Construct