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plasmid plex307 dpp4 puro  (Addgene inc)


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    Structured Review

    Addgene inc plasmid plex307 dpp4 puro
    Plasmid Plex307 Dpp4 Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plex307+dpp4+puro/pLEX307-DPP4-puro+(Plasmid+%23158451)/pmc12326988-40-12-22
    Average 93 stars, based on 15 article reviews
    plasmid plex307 dpp4 puro - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Clone Assay:

    Article Title: The KDM6A-KMT2D-p300 axis regulates susceptibility to diverse coronaviruses by mediating viral receptor expression
    Article Snippet: .. KDM6A KO clones were transduced with pLenti-hACE2-Puro (Addgene: #155295) and pLEX307-DPP4-Puro (Addgene: #158451) to overexpress human ACE2 and human DPP4, respectively. ..

    Article Title: DYRK1A promotes viral entry of highly pathogenic human coronaviruses in a kinase-independent manner.
    Article Snippet: .. For lentiviral transduction, Vero-E6 or Calu-3 cells were transduced at 50% confluency and selected with 5 μg/mL (Vero-E6) or 2.5 μg/mL (Calu-3) puromycin 48 hours later. hACE2 and hDPP4 overexpressing lines were generated by stable lentiviral delivery of pLV-EF1a-ACE2-puro (gift of Akiko Iwasaki) and pLEX307-DPP4-puro (Addgene #158451) into DYRK1A KO clones. .. Viral stocks were generated in Vero-E6 or Vero-E6-ACE2-TMPRSS2 cells seeded at approximately 80% confluency inoculated with HKU5-SARS-CoV-1-S (NR-48814), SARS-CoV-2 isolate USA-WA1/2020 (NR-52281), or MERS-CoV (icMERS-CoV EMC/2012) (NR-48813) from BEI resources at an MOI of approximately 0.01 for 3 days to generate a P1 stock.

    Article Title: The KDM6A-KMT2D-p300 axis regulates susceptibility to diverse coronaviruses by mediating viral receptor expression.
    Article Snippet: .. KDM6A KO clones were transduced with pLenti-hACE2-Puro (Addgene: #155295) and pLEX307-DPP4-Puro (Addgene: #158451) to overexpress human ACE2 and human DPP4, respectively. ..

    Transduction:

    Article Title: The KDM6A-KMT2D-p300 axis regulates susceptibility to diverse coronaviruses by mediating viral receptor expression
    Article Snippet: .. KDM6A KO clones were transduced with pLenti-hACE2-Puro (Addgene: #155295) and pLEX307-DPP4-Puro (Addgene: #158451) to overexpress human ACE2 and human DPP4, respectively. ..

    Article Title: DYRK1A promotes viral entry of highly pathogenic human coronaviruses in a kinase-independent manner.
    Article Snippet: .. For lentiviral transduction, Vero-E6 or Calu-3 cells were transduced at 50% confluency and selected with 5 μg/mL (Vero-E6) or 2.5 μg/mL (Calu-3) puromycin 48 hours later. hACE2 and hDPP4 overexpressing lines were generated by stable lentiviral delivery of pLV-EF1a-ACE2-puro (gift of Akiko Iwasaki) and pLEX307-DPP4-puro (Addgene #158451) into DYRK1A KO clones. .. Viral stocks were generated in Vero-E6 or Vero-E6-ACE2-TMPRSS2 cells seeded at approximately 80% confluency inoculated with HKU5-SARS-CoV-1-S (NR-48814), SARS-CoV-2 isolate USA-WA1/2020 (NR-52281), or MERS-CoV (icMERS-CoV EMC/2012) (NR-48813) from BEI resources at an MOI of approximately 0.01 for 3 days to generate a P1 stock.

    Article Title: The KDM6A-KMT2D-p300 axis regulates susceptibility to diverse coronaviruses by mediating viral receptor expression.
    Article Snippet: .. KDM6A KO clones were transduced with pLenti-hACE2-Puro (Addgene: #155295) and pLEX307-DPP4-Puro (Addgene: #158451) to overexpress human ACE2 and human DPP4, respectively. ..

    Expressing:

    Article Title: A trivalent protein-based pan- Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced Hela cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451 ; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM l -glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Article Title: A trivalent protein-based pan-Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses.
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. Generation of a DPP4 expressing Hela cell line To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infectionwithMERS spikepseudovirions,we transducedHela cellswith a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’smediumsupplementedwith10%heat-inactivated fetal bovine serum, 2mM L-glutamine, 1mM sodium pyruvate, 100U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Infection:

    Article Title: A trivalent protein-based pan- Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced Hela cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451 ; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM l -glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Article Title: Optimization of Cellular Transduction by the HIV-Based Pseudovirus Platform with Pan-Coronavirus Spike Proteins
    Article Snippet: HEK-ACE2 was obtained through BEI Resources, an NIH-supported program managed by the ATCC: Human Embryonic Kidney Cells (HEK-293T) Expressing Human Angiotensin-Converting Enzyme 2 (HEK-293T-hACE2 Cell Line, NR-52511). .. To generate a human DPP4 (CD26)-expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced HeLa cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez and Sho Iketani (Addgene (Watertown, MA, USA) plasmid #158451; http://n2t.net/addgene:158451 (6 July 2022); RRID:Addgene_158451)) and selected for vector-expressing cells using 1 μg/mL puromycin (MilleporeSigma (Burlington, MA, USA), P4512). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend (San Diego, CA, USA), 302705) and found to be approximately 90% DPP4-positive.

    Plasmid Preparation:

    Article Title: A trivalent protein-based pan- Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced Hela cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451 ; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM l -glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Article Title: Optimization of Cellular Transduction by the HIV-Based Pseudovirus Platform with Pan-Coronavirus Spike Proteins
    Article Snippet: HEK-ACE2 was obtained through BEI Resources, an NIH-supported program managed by the ATCC: Human Embryonic Kidney Cells (HEK-293T) Expressing Human Angiotensin-Converting Enzyme 2 (HEK-293T-hACE2 Cell Line, NR-52511). .. To generate a human DPP4 (CD26)-expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced HeLa cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez and Sho Iketani (Addgene (Watertown, MA, USA) plasmid #158451; http://n2t.net/addgene:158451 (6 July 2022); RRID:Addgene_158451)) and selected for vector-expressing cells using 1 μg/mL puromycin (MilleporeSigma (Burlington, MA, USA), P4512). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend (San Diego, CA, USA), 302705) and found to be approximately 90% DPP4-positive.

    Article Title: A trivalent protein-based pan-Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses.
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. Generation of a DPP4 expressing Hela cell line To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infectionwithMERS spikepseudovirions,we transducedHela cellswith a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’smediumsupplementedwith10%heat-inactivated fetal bovine serum, 2mM L-glutamine, 1mM sodium pyruvate, 100U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Modification:

    Article Title: A trivalent protein-based pan- Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infection with MERS spike pseudovirions, we transduced Hela cells with a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4-puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451 ; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum, 2 mM l -glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Article Title: A trivalent protein-based pan-Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses.
    Article Snippet: Genscript synthesized and inserted the genes into the pcDNA3.1(+) vector (pcDNA3.1-CoV-RaTG13del19 and pcDNA3.1-CoVRsSHC014del19). .. Generation of a DPP4 expressing Hela cell line To generate a human DPP4 (CD26) expressing target cell line (Hela-DPP4) for infectionwithMERS spikepseudovirions,we transducedHela cellswith a DPP4 encoding lentiviral vector, pLEX307-DPP4-puro (pLEX307-DPP4puro was a gift from Alejandro Chavez & Sho Iketani (Addgene plasmid # 158451; http://n2t.net/addgene:158451; RRID: Addgene_158451)) and selected for vector expressing cells using 1 μg/ml puromycin (Millipore Sigma, P4512) in complete DMEM (high glucose Dulbecco’s modified Eagle’smediumsupplementedwith10%heat-inactivated fetal bovine serum, 2mM L-glutamine, 1mM sodium pyruvate, 100U/ml penicillin, 100 μg/ml streptomycin). .. Puromycin-resistant cells were analyzed for DPP4 (CD26) surface expression by flow cytometry on an Attune Acoustic Focusing Cytometer using a mouse anti-human CD26-PE antibody (clone BA5b, Biolegend, 302705) and were found to be ~90% DPP4 positive.

    Generated:

    Article Title: DYRK1A promotes viral entry of highly pathogenic human coronaviruses in a kinase-independent manner.
    Article Snippet: .. For lentiviral transduction, Vero-E6 or Calu-3 cells were transduced at 50% confluency and selected with 5 μg/mL (Vero-E6) or 2.5 μg/mL (Calu-3) puromycin 48 hours later. hACE2 and hDPP4 overexpressing lines were generated by stable lentiviral delivery of pLV-EF1a-ACE2-puro (gift of Akiko Iwasaki) and pLEX307-DPP4-puro (Addgene #158451) into DYRK1A KO clones. .. Viral stocks were generated in Vero-E6 or Vero-E6-ACE2-TMPRSS2 cells seeded at approximately 80% confluency inoculated with HKU5-SARS-CoV-1-S (NR-48814), SARS-CoV-2 isolate USA-WA1/2020 (NR-52281), or MERS-CoV (icMERS-CoV EMC/2012) (NR-48813) from BEI resources at an MOI of approximately 0.01 for 3 days to generate a P1 stock.

    Transfection:

    Article Title: Targeting DPP4-RBD interactions by sitagliptin and linagliptin delivers a potential host-directed therapy against pan-SARS-CoV-2 infections.
    Article Snippet: Mounting was done using fluoroshield (Sigma), and cells were visualized under a confocal laser scanning inverted microscope, Carl-Zeiss LSM980 (63×)2. .. Calu-3 cells (2.0 × 105/well) were plated onto cover glasses in 9 cm2 S. Mani et al. International Journal of Biological Macromolecules 245 (2023) 125444 well of the plates, grown overnight (to ~5 × 105 cells), and was transfected with pLEX307-DPP4-puro (Addgene # 158451) alone or in combination with pCDNA3.1-RBD construct using lipofectamine 3000 (Invitrogen) following manufacturer’s instructions. .. DPP4 knockout cells were utilised in one set to transfect the pCDNA3.1-RBD construct in order to perform the DPP4-RBD immunoprecipitation assay, which would verify the absence of any experimental artefact.

    Article Title: Targeting DPP4-RBD interactions by sitagliptin and linagliptin delivers a potential host-directed therapy against pan-SARS-CoV-2 infections
    Article Snippet: Mounting was done using fluoroshield (Sigma), and cells were visualized under a confocal laser scanning inverted microscope, Carl-Zeiss LSM980 (63×)2. .. Calu-3 cells (2.0 × 10 5 /well) were plated onto cover glasses in 9 cm 2 well of the plates, grown overnight (to ∼5 × 10 5 cells), and was transfected with pLEX307-DPP4-puro (Addgene # 158451) alone or in combination with pCDNA3.1-RBD construct using lipofectamine 3000 (Invitrogen) following manufacturer's instructions. .. DPP4 knockout cells were utilised in one set to transfect the pCDNA3.1-RBD construct in order to perform the DPP4-RBD immunoprecipitation assay, which would verify the absence of any experimental artefact.

    Construct:

    Article Title: Targeting DPP4-RBD interactions by sitagliptin and linagliptin delivers a potential host-directed therapy against pan-SARS-CoV-2 infections.
    Article Snippet: Mounting was done using fluoroshield (Sigma), and cells were visualized under a confocal laser scanning inverted microscope, Carl-Zeiss LSM980 (63×)2. .. Calu-3 cells (2.0 × 105/well) were plated onto cover glasses in 9 cm2 S. Mani et al. International Journal of Biological Macromolecules 245 (2023) 125444 well of the plates, grown overnight (to ~5 × 105 cells), and was transfected with pLEX307-DPP4-puro (Addgene # 158451) alone or in combination with pCDNA3.1-RBD construct using lipofectamine 3000 (Invitrogen) following manufacturer’s instructions. .. DPP4 knockout cells were utilised in one set to transfect the pCDNA3.1-RBD construct in order to perform the DPP4-RBD immunoprecipitation assay, which would verify the absence of any experimental artefact.

    Article Title: Targeting DPP4-RBD interactions by sitagliptin and linagliptin delivers a potential host-directed therapy against pan-SARS-CoV-2 infections
    Article Snippet: Mounting was done using fluoroshield (Sigma), and cells were visualized under a confocal laser scanning inverted microscope, Carl-Zeiss LSM980 (63×)2. .. Calu-3 cells (2.0 × 10 5 /well) were plated onto cover glasses in 9 cm 2 well of the plates, grown overnight (to ∼5 × 10 5 cells), and was transfected with pLEX307-DPP4-puro (Addgene # 158451) alone or in combination with pCDNA3.1-RBD construct using lipofectamine 3000 (Invitrogen) following manufacturer's instructions. .. DPP4 knockout cells were utilised in one set to transfect the pCDNA3.1-RBD construct in order to perform the DPP4-RBD immunoprecipitation assay, which would verify the absence of any experimental artefact.



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    Figure 1. Designed and engineered high-affinity <t>DPP4</t> by direct evolution (A) 293T cells expressing each DPP4 mutant were incubated with the spike RBD of MERS-CoV labeled with superfolder GFP (sfGFP). (B) Random mutations were introduced into the DPP4 interface region (aa 264–350) by error-prone PCR amplification. The constructed DPP4 mutant plasmid library was diluted and transfected into 293T cells. Using a cell sorter, the cells incubated with RBD sfGFP were displayed for RDB binding (sfGFP) and DPP4-HA expression (Alexa 647) and the top 0.05% high-level-bound population was harvested. Extracted mRNA from the sorted cells was used for the next cycle. (C) Competitive inhibition activity was assessed in three types of high-affinity DPP4 clones. Each soluble DPP4 mutant was incubated with the RBD-sfGFP for 30 min and then the mixture was transferred to native DPP4-expressing 293T cells. Inhibition rate was calculated from the number of sfGFP-binding cells using flow cytometry. n = 3 technical replicates. (D) Neutralizing activity of Fc-DPP4 was evaluated against MERS-CoV pseudovirus. Fc-DPP4 is human IgG1 Fc fused N-terminal to DPP4 mutants. n = 4.
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    Image Search Results


    Figure 1. Designed and engineered high-affinity DPP4 by direct evolution (A) 293T cells expressing each DPP4 mutant were incubated with the spike RBD of MERS-CoV labeled with superfolder GFP (sfGFP). (B) Random mutations were introduced into the DPP4 interface region (aa 264–350) by error-prone PCR amplification. The constructed DPP4 mutant plasmid library was diluted and transfected into 293T cells. Using a cell sorter, the cells incubated with RBD sfGFP were displayed for RDB binding (sfGFP) and DPP4-HA expression (Alexa 647) and the top 0.05% high-level-bound population was harvested. Extracted mRNA from the sorted cells was used for the next cycle. (C) Competitive inhibition activity was assessed in three types of high-affinity DPP4 clones. Each soluble DPP4 mutant was incubated with the RBD-sfGFP for 30 min and then the mixture was transferred to native DPP4-expressing 293T cells. Inhibition rate was calculated from the number of sfGFP-binding cells using flow cytometry. n = 3 technical replicates. (D) Neutralizing activity of Fc-DPP4 was evaluated against MERS-CoV pseudovirus. Fc-DPP4 is human IgG1 Fc fused N-terminal to DPP4 mutants. n = 4.

    Journal: Cell Biomaterials

    Article Title: Engineered DPP4 decoy confers broad-spectrum inhibition of MERS-CoV infection

    doi: 10.1016/j.celbio.2025.100018

    Figure Lengend Snippet: Figure 1. Designed and engineered high-affinity DPP4 by direct evolution (A) 293T cells expressing each DPP4 mutant were incubated with the spike RBD of MERS-CoV labeled with superfolder GFP (sfGFP). (B) Random mutations were introduced into the DPP4 interface region (aa 264–350) by error-prone PCR amplification. The constructed DPP4 mutant plasmid library was diluted and transfected into 293T cells. Using a cell sorter, the cells incubated with RBD sfGFP were displayed for RDB binding (sfGFP) and DPP4-HA expression (Alexa 647) and the top 0.05% high-level-bound population was harvested. Extracted mRNA from the sorted cells was used for the next cycle. (C) Competitive inhibition activity was assessed in three types of high-affinity DPP4 clones. Each soluble DPP4 mutant was incubated with the RBD-sfGFP for 30 min and then the mixture was transferred to native DPP4-expressing 293T cells. Inhibition rate was calculated from the number of sfGFP-binding cells using flow cytometry. n = 3 technical replicates. (D) Neutralizing activity of Fc-DPP4 was evaluated against MERS-CoV pseudovirus. Fc-DPP4 is human IgG1 Fc fused N-terminal to DPP4 mutants. n = 4.

    Article Snippet: Human DPP4 (Addgene #158451) was cloned into the KpnI-XhoI sites of pcDNA4TO (Thermo Fisher Scientific) with a C-terminal HA tag.

    Techniques: Expressing, Mutagenesis, Incubation, Labeling, Construct, Plasmid Preparation, Transfection, Binding Assay, Inhibition, Activity Assay, Clone Assay, Cytometry

    Figure 6. Engineered DPP4 decoy exhibited prophylactic efficacy against MERS-CoV infection (A) Schematic of the mouse experiment. 3 h prior to MERS-CoV infection with 1 3 105 plaque-forming units, hDPP4 transgenic mice were intranasally admin- istered 200 mg of the engineered DPP4 decoy. Lung tissues were harvested 1 and 3 days after infection. (B) Genomic MERS-CoV RNA and subgenomic RNA extracted from harvested lung tissues were evaluated as copies per mg of cellular transcripts. Viral titration of lung tissues was performed using the TCID50 assay. Data are expressed as mean ± SD (n = 4 per group). p values were determined using two-way ANOVA with Sida´ k’s multiple comparison test. Dashed lines indicate the detection limit. (C) HE staining and immunohistochemistry of MERS-CoV NP and CD3. Scale bars, 100 mm. (D) MERS-CoV NP- and CD3-positive cells were quantified as densities per mm2. For each individual, the density of three lobules (left, right cranial, and right middle) was quantified and averaged. Data are expressed as mean ± SD (n = 4 per group). p values were determined by unpaired t test.

    Journal: Cell Biomaterials

    Article Title: Engineered DPP4 decoy confers broad-spectrum inhibition of MERS-CoV infection

    doi: 10.1016/j.celbio.2025.100018

    Figure Lengend Snippet: Figure 6. Engineered DPP4 decoy exhibited prophylactic efficacy against MERS-CoV infection (A) Schematic of the mouse experiment. 3 h prior to MERS-CoV infection with 1 3 105 plaque-forming units, hDPP4 transgenic mice were intranasally admin- istered 200 mg of the engineered DPP4 decoy. Lung tissues were harvested 1 and 3 days after infection. (B) Genomic MERS-CoV RNA and subgenomic RNA extracted from harvested lung tissues were evaluated as copies per mg of cellular transcripts. Viral titration of lung tissues was performed using the TCID50 assay. Data are expressed as mean ± SD (n = 4 per group). p values were determined using two-way ANOVA with Sida´ k’s multiple comparison test. Dashed lines indicate the detection limit. (C) HE staining and immunohistochemistry of MERS-CoV NP and CD3. Scale bars, 100 mm. (D) MERS-CoV NP- and CD3-positive cells were quantified as densities per mm2. For each individual, the density of three lobules (left, right cranial, and right middle) was quantified and averaged. Data are expressed as mean ± SD (n = 4 per group). p values were determined by unpaired t test.

    Article Snippet: Human DPP4 (Addgene #158451) was cloned into the KpnI-XhoI sites of pcDNA4TO (Thermo Fisher Scientific) with a C-terminal HA tag.

    Techniques: Infection, Transgenic Assay, Titration, TCID50 Assay, Comparison, Staining, Immunohistochemistry