Plasmid Preparation:Article Title: Cyclophilin J Reprograms Tumor-associated Macrophages to Exert an Anti-tumor Effect in Liver Cancer
Article Snippet: Additionally, when BMDMs are infected with recombinant adeno-associated virus (rAAV), the Raji or K562 cells are then labeled with CellTraceTM Far Red (APC channel, Thermo Fisher Scientific, # C34572 , USA), and all cells were collected and stained with PE-CD11b. .. The triple plasmid system (the vector pscAAV-CAG-GFP (Addgene, #83279) or pscAAV-CAG-CYPJ or pscAAV-CAG-mutCYPJ, the serotype AAV1 Rep/Cap (Addgene, #112862) and helper plasmid (Addgene, #112867) were used for rAAV production. ..
Article Title: A photo-cross-linking approach to monitor protein dynamics in living cells.
Article Snippet: Accepted Manuscript A photo-cross-linking approach to monitor protein dynamics in living cells Ryoji Miyazaki, Yoshinori Akiyama, Hiroyuki Mori PII: S0304-4165(19)30053-4 DOI: https://doi.org/10.1016/j.bbagen.2019.03.003 Reference: BBAGEN 29317 To appear in: BBA - General Subjects Received date: 20 December 2018 Revised date: 26 February 2019 Accepted date: 4 March 2019 Please cite this article as: R. Miyazaki, Y. Akiyama and H. Mori, A photo-cross-linking approach to monitor protein dynamics in living cells, BBA - General Subjects, https://doi.org/10.1016/j.bbagen.2019.03.003 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form.
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [23], SIVCPZ, SIVAGM−SAB, or SIVMAC CA (gifts from Theodora Hatziioannou) [24]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [57] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [58]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [ ], SIV CPZ , SIV AGM-SAB , or SIV MAC CA (gifts from Theodora Hatziioannou) [ ]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [ ] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [ ]. ..
Article Title: Optimization of T4 phage engineering via CRISPR/Cas9.
Article Snippet: This group designed and employed the DS-SPCas (Addgene no. 48645) and the PM-SP!TB (Addgene no. 48650) plasmids in which Cas9 and the crRNA exist on separate plasmids27,36. .. Finally, we evaluated the dual plasmid system introduced by Marraffini et al. consisting of pCas9 (Addgene no.42876) and pCRISPR (Addgene no. 42875)37. ..
Construct:Article Title: A photo-cross-linking approach to monitor protein dynamics in living cells.
Article Snippet: Accepted Manuscript A photo-cross-linking approach to monitor protein dynamics in living cells Ryoji Miyazaki, Yoshinori Akiyama, Hiroyuki Mori PII: S0304-4165(19)30053-4 DOI: https://doi.org/10.1016/j.bbagen.2019.03.003 Reference: BBAGEN 29317 To appear in: BBA - General Subjects Received date: 20 December 2018 Revised date: 26 February 2019 Accepted date: 4 March 2019 Please cite this article as: R. Miyazaki, Y. Akiyama and H. Mori, A photo-cross-linking approach to monitor protein dynamics in living cells, BBA - General Subjects, https://doi.org/10.1016/j.bbagen.2019.03.003 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form.
In Vivo:Article Title: A photo-cross-linking approach to monitor protein dynamics in living cells.
Article Snippet: Accepted Manuscript A photo-cross-linking approach to monitor protein dynamics in living cells Ryoji Miyazaki, Yoshinori Akiyama, Hiroyuki Mori PII: S0304-4165(19)30053-4 DOI: https://doi.org/10.1016/j.bbagen.2019.03.003 Reference: BBAGEN 29317 To appear in: BBA - General Subjects Received date: 20 December 2018 Revised date: 26 February 2019 Accepted date: 4 March 2019 Please cite this article as: R. Miyazaki, Y. Akiyama and H. Mori, A photo-cross-linking approach to monitor protein dynamics in living cells, BBA - General Subjects, https://doi.org/10.1016/j.bbagen.2019.03.003 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form.
Generated:Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [23], SIVCPZ, SIVAGM−SAB, or SIVMAC CA (gifts from Theodora Hatziioannou) [24]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [57] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [58]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [ ], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [ ] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [ ], L-VSV-G for expression of VSV-G envelope [ ], CMV-tat for transactivation [ ], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [ ]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [ ], SIV CPZ , SIV AGM-SAB , or SIV MAC CA (gifts from Theodora Hatziioannou) [ ]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [ ] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [ ]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [40], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [22] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [59], L-VSV-G for expression of VSV-G envelope [59], CMV-tat for transactivation [59], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [60]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
Transfection:Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [23], SIVCPZ, SIVAGM−SAB, or SIVMAC CA (gifts from Theodora Hatziioannou) [24]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [57] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [58]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [ ], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [ ] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [ ], L-VSV-G for expression of VSV-G envelope [ ], CMV-tat for transactivation [ ], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [ ]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [ ], SIV CPZ , SIV AGM-SAB , or SIV MAC CA (gifts from Theodora Hatziioannou) [ ]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [ ] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [ ]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [40], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [22] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [59], L-VSV-G for expression of VSV-G envelope [59], CMV-tat for transactivation [59], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [60]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
Expressing:Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [23], SIVCPZ, SIVAGM−SAB, or SIVMAC CA (gifts from Theodora Hatziioannou) [24]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [57] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [58]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [ ], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [ ] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [ ], L-VSV-G for expression of VSV-G envelope [ ], CMV-tat for transactivation [ ], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [ ]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor
Article Snippet: .. Fluorescent reporter viruses were generated by transfection of a three-plasmid system: pMD2.G (Addgene, Watertown, MA, #12259, a gift from Didier Trono) for expression of VSV-G envelope; a variable plasmid for expression of a chimeric gag/pol containing an NL4-3 backbone and HIV-1 [ ], SIV CPZ , SIV AGM-SAB , or SIV MAC CA (gifts from Theodora Hatziioannou) [ ]; and either pALPS-eGFP (Addgene #101323, a gift from Jeremy Luban) [ ] or pHIV-zsGreen (Addgene #18121, a gift from Bryan Welm and Zena Werb) [ ]. ..
Article Title: Primate TRIM34 is a broadly-acting, TRIM5-dependent lentiviral restriction factor.
Article Snippet: .. N-MLV reporter viruses were generated by transfection of a three-plasmid system: pCIG3-N for expression N-MLV gag/pol (Addgene #132941, a gift from Jeremy Luban) [40], pQCXIP-eGFP (a gift from Jeannette Tenthorey) [22] for expression of a fluorescent reporter, and pMD2.G for expression of VSV-G envelope. .. NB-MLV reporter viruses were generated by transfection of a four-plasmid system: JK3 for expression of NB-MLV gag/pol [59], L-VSV-G for expression of VSV-G envelope [59], CMV-tat for transactivation [59], and pQCXIP-eGFP for expression of a fluorescent reporter. pLentiCRISPR-v2 (Addgene #52961, a gift from Feng Zhang) constructs were generated by BsmBI (New England BioLabs, Ipswich, MA, #R0580) restriction cloning of TRIM5 guides into pLentiCRISPR-v2, a lentiviral vector encoding for Cas9 [60]. pLentiCRISPR-v2 containing guides were transfected with pSPAX2 (Addgene #12260, a gift from Didier Trono) and pMD2.G to generate lentiviral particles.
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