Plenti vector

Manufactured by OriGene

The PLenti vector is a lentiviral expression vector designed for the stable transduction and expression of genes of interest in mammalian cells. It contains the necessary elements for the production of replication-incompetent lentiviral particles, enabling the efficient delivery and integration of the target gene into the host cell genome.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Flag m2, by Merck Group (1 mentions) Las 3000 luminescent image analyzer system, by Fujifilm (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Amicon ultra 15 centrifugal filter unit, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) Lenti x gostix plus kit, by Takara Bio (1 mentions) Dmem 5 mm glucose, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) H1975, by American Type Culture Collection (1 mentions)

Spelling variants of this product

PLenti-C-Myc-DDK vector (6 citations) PLenti-C-Myc-DDK-IRES-Puro vector (6 citations) PLenti-C-mGFP-P2A-Puro vector (5 citations) PLenti-C-Myc-DDK-IRES-Neo vector (3 citations) PLenti-C-Myc-DDK-IRES-Puro (PCMV) vector (3 citations) PLenti-DDK-P2A-Puro empty vector (3 citations) PLenti-Guide-Puro- Crispr vector (2 citations) PLenti-C-Myc-DDK-IRES-Puro expression vector (2 citations) PLenti-C-mGFP-P2A-Puro Lentiviral Gene Expression Vector (2 citations) PLenti S100b vector (2 citations)

4 protocols using plenti vector

Generation of ADAR1 Catalytic Mutant

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To generate the editing deficient ADAR1 construct, site-directed mutagenesis was performed using the pLenti-ADAR1 construct (purchased by Origene) to integrate the point mutation at aa912 which leads to the substitution of glutamate (E, GAA) with alanine (A, GCA), previously shown to inactivate the catalytic activity of ADAR1 [28 (link),29 (link)]. HUVECs at 80–90% confluency were transfected with 5 μg of either pLenti-ADAR1, pLenti-ADAR1 E/A (generated editing deficient mutant) or an empty pLenti vector (control, purchased also by Origene). Transient plasmid transfection was performed with Lipofectamine 2000 (Invitrogen) according to the manufacturers protocol. After 48 h of incubation at 37 °C, 5% CO₂ cells were harvested for RNA expression studies, as described below.

Vlachogiannis N.I., Sachse M., Georgiopoulos G., Zormpas E., Bampatsias D., Delialis D., Bonini F., Galyfos G., Sigala F., Stamatelopoulos K., Gatsiou A, & Stellos K. (2021). Adenosine-to-inosine Alu RNA editing controls the stability of the pro-inflammatory long noncoding RNA NEAT1 in atherosclerotic cardiovascular disease. Journal of Molecular and Cellular Cardiology, 160, 111-120.

+ Open protocol

+ Expand

Characterization of ERCC6 and ERCC4 Deficient Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

ERCC6 (GM16095)- and ERCC4 (GM08437)-deficient immortalized fibroblast cell lines (Coriell Cell Repository) were cultured in DMEM (Invitrogen), supplemented with 10% FBS, 1% Penicillin Streptomycin and L-glutamine. ERCC6-deficient line was complemented with either wild-type or mutant N-terminal Flag-tagged ERCC6, cloned in a pOZ vector. The ERCC4-deficient line was complemented with C-terminal Myc-tagged wild-type or mutant ERCC4, cloned in a pLenti vector (Origene).
For Western blotting, whole cell extracts were prepared by lysing cells in RIPA with complete protease inhibitor. Lysates were resolved on a polyacrylamide gel, transferred to a PVDF membrane, and incubated with primary antibodies [BRCA2 (ab-1; Calbiochem), BRCA1 (ab-1; Calbiochem), ERCC4 (D3G8C; Cell Signaling Technology), ERCC6 (D-7; Santa Cruz Biotechnology), Flag (M2; Sigma), and β-actin (Cell Signaling Technology)]. Signal was detected using an ECL kit (Pierce) and visualized with a Fuji LAS-3000 luminescent image analyzer system.

Ceccaldi R., O'Connor K.W., Mouw K.W., Li A.Y., Matulonis U.A., D'Andrea A.D, & Konstantinopoulos P.A. (2015). A Unique Subset of Epithelial Ovarian Cancers with Platinum Sensitivity and PARP Inhibitor Resistance. Cancer research, 75(4), 628-634.

+ Open protocol

+ Expand

Lentiviral Transduction of Nasal Organoids

Check if the same lab product or an alternative is used in the 5 most similar protocols

The FLAG-tagged spike genes of WT, B.1.1.529, and BA.5 were inserted into a pLenti vector (Origene, PS100069). Lenti-X 293T cells seeded in a 6-well plate were cotransfected with three plasmids, including 12 μg pLenti-spike, 6 μg pspax2, and 6 μg pmd2g, using Lipofectamine 3000 (ThermoFisher). The culture media containing lentivirus particles were harvested 48 h posttransfection. After concentration with Amicon Ultra-15 Centrifugal Filter Unit (Millipore, UFC910008), we titrated the lentiviruses using a Lenti-X GoStix Plus kit (Takara Bio, 631280). Nasal organoid monolayers were transduced with lentiviruses at 5 MOI with 10 μg/mL polybrene. The transduced nasal organoids were fixed with 4% PFA at 48 h posttransduction and applied to immunostaining using an anti-FLAG (Sigma, F7425), followed by confocal imaging as described above (36 (link)).

Li C., Huang J., Yu Y., Wan Z., Chiu M.C., Liu X., Zhang S., Cai J.P., Chu H., Li G., Chan J.F., To K.K., Yang Z., Jiang S., Yuen K.Y., Clevers H, & Zhou J. (2023). Human airway and nasal organoids reveal escalating replicative fitness of SARS-CoV-2 emerging variants. Proceedings of the National Academy of Sciences of the United States of America, 120(17), e2300376120.

+ Open protocol

+ Expand

Overexpressing EIF3F in Human Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

A549, HeLa, H460, and H1975 human cells were obtained from ATCC (Bethesda, MD, USA). Cell culture was performed in DMEM 5 mM glucose (Gibco), supplemented with 10% FBS (Gibco) and penicillin–streptomycin 1× (100×; Gibco). The lentiviral vector used for human EIF3F overexpression was constructed by inserting EIF3F cDNA into pLenti vector, which contains a FLAG tag and a gene for puromycin resistance (OriGene). Lentiviral particles were produced by transient transfection of 293T cells using a calcium phosphate transfection technique. Prior to infection the medium was removed and 5 × 10⁵ cells were incubated with viral supernatants for 48 h at 37 °C in the presence of 8 μg/ml of protamine sulfate. EIF3F overexpressing cells were selected using promycin treatment (1 µg/mL) in culture medium.

Esteves P., Dard L., Brillac A., Hubert C., Sarlak S., Rousseau B., Dumon E., Izotte J., Bonneu M., Lacombe D., Dupuy J.W., Amoedo N, & Rossignol R. (2019). Nuclear control of lung cancer cells migration, invasion and bioenergetics by eukaryotic translation initiation factor 3F. Oncogene, 39(3), 617-636.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!