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nhei vp64 pmei fragment  (Addgene inc)


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    Structured Review

    Addgene inc nhei vp64 pmei fragment
    Nhei Vp64 Pmei Fragment, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/vp64+fragment/pcDNA3%2E1-Cry2FL-VP64+(Plasmid+%2360554)/bio_rxiv__2025__11__13__688218-152-1-6
    Average 92 stars, based on 5 article reviews
    nhei vp64 pmei fragment - by Bioz Stars, 2026-10
    92/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
    Article Snippet: The GB1 fragment was synthesized from Genewiz. .. The VP64 fragment was PCR amplified from the NMnCas9-VP64 (Addgene number 48676) using primer pVP64F and pVP64R. pSA041 (CAG-NLS-ScFV-GB1-FKBP) was made in a Golden Gate reaction with ScFV fragment, GB1 fragment, FKBP fragment and the pA017 vector. ..

    Article Title: Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
    Article Snippet: The FRB* was synthesized from Genewiz. .. The VP64 fragment was PCR amplified from the NMnCas9-VP64 (Addgene number 48676) using primer pVP64F and pVP64R. pSA044 (PENTR-L1-NLS-LacZ-2A-TALE14-4xTarget^miR18a-L2) was made by inserting the LacZ fragment into the pWX45 (pENTR-L1-mkate2-T2A-TALE144xTarget^miR18a-L2)1 by using MfeI and PspOM1.The LacZ fragment was PCR amplified from pDS001 using primer M13F and pLacZR. pSA045 (pENTR-L1-NLS-LacZ-NLS-T2A-TALE14-4xTarget^FF4-L2) was made by inserting the LacZ fragment into the pWX22 (pENTR-L1-dTA-T2A-TALER144xTarget^FF4-L2)1 by using MfeI and PspOMI. ..

    Amplification:

    Article Title: Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
    Article Snippet: The GB1 fragment was synthesized from Genewiz. .. The VP64 fragment was PCR amplified from the NMnCas9-VP64 (Addgene number 48676) using primer pVP64F and pVP64R. pSA041 (CAG-NLS-ScFV-GB1-FKBP) was made in a Golden Gate reaction with ScFV fragment, GB1 fragment, FKBP fragment and the pA017 vector. ..

    Article Title: Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
    Article Snippet: The FRB* was synthesized from Genewiz. .. The VP64 fragment was PCR amplified from the NMnCas9-VP64 (Addgene number 48676) using primer pVP64F and pVP64R. pSA044 (PENTR-L1-NLS-LacZ-2A-TALE14-4xTarget^miR18a-L2) was made by inserting the LacZ fragment into the pWX45 (pENTR-L1-mkate2-T2A-TALE144xTarget^miR18a-L2)1 by using MfeI and PspOM1.The LacZ fragment was PCR amplified from pDS001 using primer M13F and pLacZR. pSA045 (pENTR-L1-NLS-LacZ-NLS-T2A-TALE14-4xTarget^FF4-L2) was made by inserting the LacZ fragment into the pWX22 (pENTR-L1-dTA-T2A-TALER144xTarget^FF4-L2)1 by using MfeI and PspOMI. ..

    Plasmid Preparation:

    Article Title: Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
    Article Snippet: The GB1 fragment was synthesized from Genewiz. .. The VP64 fragment was PCR amplified from the NMnCas9-VP64 (Addgene number 48676) using primer pVP64F and pVP64R. pSA041 (CAG-NLS-ScFV-GB1-FKBP) was made in a Golden Gate reaction with ScFV fragment, GB1 fragment, FKBP fragment and the pA017 vector. ..

    Retroviral:

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses.
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (SigmaAldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector84 and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome (gRandom) (Supplementary Table 1 within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (Sigma-Aldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1- NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome ( gRandom ) (Supplementary Table within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).

    Clone Assay:

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses.
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (SigmaAldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector84 and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome (gRandom) (Supplementary Table 1 within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (Sigma-Aldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1- NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome ( gRandom ) (Supplementary Table within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).

    Cloning:

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses.
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (SigmaAldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector84 and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome (gRandom) (Supplementary Table 1 within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).

    Article Title: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses
    Article Snippet: Naïve B cells were activated by addition of 5-25 μg/ml LPS (Sigma-Aldrich) and 5 ng/ml of mouse recombinant IL-4 (Sigma-Aldrich) (L-I), or 5 μg/ml LPS, 10 ng/ml BAFF (PeproTech) and 2 ng/ml TGFβ (L-B-T), or 5 μg/ml LPS only (L). .. The retroviral pMSCV-gRNA-scfv-VP64-eGFP vector was built by combining the scfv fragment obtained from the phr-scfvGCN4-sfGFP-GB1- NLS-dWPRE vector (addgene #60906) and the VP64 fragment from the PB-TRE-dCas9-VPR vector (addgene #63800) into the MSCV_hU6_CcdB_PGK_Puro_T2A_BFP vector and BFP was replaced with eGFP. gRNAs against the promoter regions of Rif1 and Prdm1 as well as gRNAs targeting regions not present in the mouse genome ( gRandom ) (Supplementary Table within Supplementary file pdf) were cloned into pMSCV-gRNA-scfv-VP64 using the two BbsI cloning sites. .. Individual gRNA-containing constructs were transfected together with pCL-Eco into the HEK293T derivative cell line BOSC23 using FuGENE HD Transfection Reagent (Promega).



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