Pentr vector

Manufactured by Addgene

Sourced in United States

The pENTR vector is a plasmid designed for Gateway cloning. It is used to create entry clones that can then be recombined with destination vectors to generate expression constructs. The pENTR vector contains attL1 and attL2 recombination sites, which allow for the transfer of DNA sequences between entry and destination vectors using the Gateway cloning system.

Automatically generated - may contain errors

Lab products found in correlation

Snail1, by Thermo Fisher Scientific (2 mentions) Plenti6 vector, by Addgene (2 mentions) Shrna clones for ddr2, by Thermo Fisher Scientific (2 mentions) Trans lentiviral packaging system, by Addgene (2 mentions) Lentivirus expression system, by Thermo Fisher Scientific (2 mentions) Blasticidin, by InvivoGen (2 mentions) Puromycin, by InvivoGen (2 mentions) Puromycin, by Merck Group (1 mentions) Twist1, by Thermo Fisher Scientific (1 mentions)

3 protocols using pentr vector

Lentiviral Knockdown of SUMO1P3

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For lentiviral construction, the sequence of short hairpin RNA (shRNA) targeting SUMO1P3 (shSUMO1P3) or a scrambled shRNA (shNC) was cloned into pENTR vector (Addgene, Cambridge, MA, USA). pMDLg/pRRE and pRSV-REV are lentiviral packaging plasmids. HEK293T cells were cotransfected with the recombinant pENTR, pMDLg/pRRE and pRSV-REV plasmids. After 48 h, the medium was collected for the purification of viruses. MHCC97H-luciferase (MHCC97H-luc) cells were incubated with virus-containing supernatants in the presence of 8 mg/ml polybrene. The stable-infected cells were screened for 2 weeks by 5 μg/mL of puromycin (Sigma).

Hu S., Liu J., Feng S., Wang Y, & Liu H. (2021). LncRNA SUMO1P3 acts as a prognostic biomarker and promotes hepatocellular carcinoma growth and metastasis. Aging (Albany NY), 13(9), 12479-12492.

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Overexpression and Knockdown of EMT Regulators

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Full-length FOXQ1, SNAIL1, TWIST1, ZEB2, and DDR2 plasmids were purchased from Open Biosystems. These genes were subcloned into the pENTR vector (RRID:Addgene_149548) and transferred into a pLenti6 vector (BRID: Addgene_21691) via homologous recombination. The lentivirus for the full-length gene was then generated using the lentivirus-expression system (Invitrogen). In addition, a set of short hairpin RNA (shRNA) clones for DDR2, FOXQ1, or SNAIL1 was purchased from Open Biosystems. The information for the effective shRNA is available in Supplementary Primers and shRNA information. The lentivirus for the shRNA was then generated using the Trans-Lentiviral packaging system (Addgene). The generated lentivirus was then used to infect the targeted model cells. Stable cells were generated after being selected with Blasticidin (10 μg /mL) for the overexpression model or puromycin (12 μg/mL; Invivogen) for the knockdown model.

Mitchell A.V., Wu J., Meng F., Dong L., Block C.J., Song W.M., Zhang B., Li J, & Wu G. (2022). DDR2 Coordinates EMT and Metabolic Reprogramming as a Shared Effector of FOXQ1 and SNAI1. Cancer Research Communications, 2(11), 1388-1403.

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Overexpression and Knockdown of EMT Regulators

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Full-length FOXQ1, SNAIL1, TWIST1, ZEB2, and DDR2 plasmids were
purchased from Open Biosystems (Huntsville, AL). These genes were sub-cloned
into the pENTR vector (RRID:Addgene_149548) and transferred into a pLenti6
vector (BRID: Addgene_21691) via homologous recombination. The lentivirus for
the full-length gene was then generated using the lentivirus-expression system
(Invitrogen, Carlsbad, CA). In addition, a set of shRNA clones for DDR2, FOXQ1,
or SNAIL1 was purchased from Open Biosystems. The information for the effective
shRNA is available in Supplementary Primers and shRNA information. The lentivirus for the
shRNA was then generated using the Trans-Lentiviral packaging system (Addgene,
Watertown, MA). The generated lentivirus was then used to infect the targeted
model cells. Stable cells were generated after being selected with Blasticidin
(10 μg /ml) for the overexpression model or puromycin (12 μg /ml)
(Invivogen, San Diego, CA) for the knockdown model.

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