The lentiviral CRISPR/Cas9-mediated BAG3 gene-editing vectors were constructed by annealing gRNA oligonucleotide pairs and subcloning them into a pLenti-Cas9-sgRNA-puro lentivirus vector (GeneChem Co., Ltd.). Genes encoding BAG3 and IMP3 were cloned into a pGC-LV-GV166 lentivirus vector (GeneChem Co., Ltd.). The shRNA targets Ago2, Roquin, IMP1, IMP3, and HSP70 were designed and cloned into a GV118 lentivirus vector (GeneChem Co., Ltd.). DNA sequencing was performed by GeneChem Co., Ltd. to verify the sequence of the insert, and the identities were 100%. After construction, the recombined lentivirus vector, the pHelper 1.0 plasmid, and the pHelper 2.0 plasmid (Invitrogen) were cotransfected into 293T cells using Lipofectamine 2000 (Invitrogen). Recombinant lentiviruses were harvested at 72 h after transfection, centrifuged to get rid of cell debris, and then filtered through a 0.22-µm cellulose acetate filter. Ultimately, a concentrated lentivirus solution was obtained with a final titer of 1.0 × 109 TU/ml.
Gv118 lentivirus vector
Manufactured by Genechem
The GV118 lentivirus vector is a genetic engineering tool used for the delivery of genetic material into target cells. It is a replication-deficient lentiviral vector that can efficiently transduce a wide range of cell types, including both dividing and non-dividing cells. The GV118 vector is designed to provide stable and long-term expression of the delivered genetic cargo within the transduced cells.
Automatically generated - may contain errors
Lab products found in correlation
Plenti cas9 sgrna puro lentivirus vector, by Genechem (3 mentions)
Pgc lv gv166 lentivirus vector, by Genechem (2 mentions)
Phelper 1.0 plasmid, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Liposome 3000, by Thermo Fisher Scientific (1 mentions)
3 protocols using gv118 lentivirus vector
1
Lentiviral CRISPR/Cas9 gene-editing vectors
2
Lentiviral CRISPR/Cas9 for PHGDH Editing
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Lentiviral CRISPR/cas9 mediated PHGDH gene editing vector was constructed by annealing gRNA oligonucleotide pairs and subcloning them into pLenti-Cas9-sgRNA-puro lentivirus vector (Genechem Co., Ltd.). The gene encoding PHGDH labeled with Myc epitope (Myc-PHGDH) and RMRP were cloned into pGCLV-GV166 lentivirus vector (Genechem Co., Ltd.). The shRNA targeting RMRP was designed and cloned into GV118 lentivirus vector (Genechem Co., Ltd.). The RNA sequence information was shown in the Supplementary Table 1
3
CRISPR/Cas9-Mediated AUF1 Gene Editing
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The lentiviral CRISPR/Cas9-mediated AUF1 gene-editing vectors were generated by annealing gRNA oligonucleotide pairs and subcloning them into a pLenti-Cas9-sgRNA-puro lentivirus vector (GeneChem Co., Ltd.). The shRNA targeting TRIM58 (shTRIM58) or ZBTB2 (shZBTB2) were designed and cloned into a GV118 lentivirus vector (GeneChem Co., Ltd.). DNA sequencing was performed to verify the sequence of the insert, and the identities were 100%. A concentrated lentivirus solution was obtained with a final titer of 1.0 × 109 TU/ml.
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