Log In Sign up
The largest database of trusted experimental protocols

Pgl3 promoter vector

Manufactured by Genechem
Visit Supplier

The PGL3 promoter vector is a plasmid used in molecular biology experiments to study gene expression. It contains the firefly luciferase gene under the control of a promoter sequence, which can be used to measure promoter activity in various cell lines.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dual luciferase reporter assay system, by Promega (1 mentions)

Close spelling variants of this product

PGL3 promoter PGL3 vector

4 protocols using pgl3 promoter vector

1

Validating miR-4516 Binding to MAPK10

Check if the same lab product or an alternative is used in the 5 most similar protocols
To validate the predicted miR-4516 binding site, a fragment of MAPK10 3’ UTR or a double mutated sequence of the predicted target sites were inserted into the XbaI site of pGL3 promoter vector, and for cis-regulation analysis, the MAPK10 3’ UTR containing either the haplotype ACT (rs1201-rs7699978-rs2575675) or GAC (rs1201-rs7699978-rs2575675) was inserted into the XbaI site of pGL3 promoter vector (Genechem, Shanghai, China). By using lipofectamine 2000 (Thermo Fisher Scientific), the constructs were co-transfected with either miR-4516 mimics, or control mimics (50 nM final concentration) to the SH-SY5Y cells. 48 h after transfections, luciferase luminescence (firefly and renilla) was measured by the Dual-Luciferase® Reporter Assay System (Promega).
Wang Y., Jiang Q., Chakravarti A., Cai H., Xu Z., Wu W., Gu B., Li L, & Cai W. (2020). MicroRNA-4516-mediated regulation of MAPK10 relies on 3’UTR cis-acting variants and contributes to the altered risk of Hirschsprung disease. Journal of medical genetics, 57(9), 634-642.
+ Open protocol
+ Expand
2

AHNAK 3'-UTR Luciferase Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
pGL3-AHNAK-WT and pGL3-AHNAK-MUT were constructed by inserting the predicted 3′-UTR sequence of AHNAK binding to miR-1251 and the mutated sequence into the pGL3 promoter vector (Genechem, Shanghai, China). For reporter assay, cells were plated into 24-well-plates and transfected with 100 ng of pGL3-AHNAK-WT and pGL3-AHNAK-MUT, 50 nM miR-1251 mimics, and negative control using Lipofectamine 2000. Renilla luciferase vector pRL-SV40 (5 ng) was transfected into cells as control. Based on the obtained ratio, the activation degree of target reporter genes in different sample was compared.
Zhou L., Zhi Z., Chen P., Du C., Wang B., Fang X., Tang W, & Li H. (2022). LncRNA-RMST Functions as a Transcriptional Co-regulator of SOX2 to Regulate miR-1251 in the Progression of Hirschsprung's Disease. Frontiers in Pediatrics, 10, 749107.
+ Open protocol
+ Expand
3

Investigating miR-1251 regulation of AHNAK

Check if the same lab product or an alternative is used in the 5 most similar protocols
The predicted 3'-UTR sequence of AHNAK binding to miR-1251 and the mutated sequence were inserted into the pGL3 promoter vector (Genechem, Shanghai, China) named pGL3-AHNAK-WT and pGL3-AHNAK-MUT. For reporter assay, cells were planted into 24-well plates and transfected with 100ng of pGL3-AHNAK-WT and pGL3-AHNAK-MUT, 50nM miR-1251 mimics and negative control using Lipofectamine 2000. Renilla luciferase vector pRL-SV40 (5 ng) was transfected into cells as control. Based on the obtained ratio, the activation degree of target reporter genes in different sample was compared.
preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this this version posted September 24, 2020. ; https://doi.org/10.1101/2020.09.23.309930 doi: bioRxiv preprint 5
Zhou L., Zhi Z., Chen P., Wei Z., Du C., Wang B., Fang X., Tang W., & Li H. (2020). lncRNA-RMST functioned as a SOX2 transcription co-regulator to regulate miR-1251 in the progression of Hirschsprung’s disease.
+ Open protocol
+ Expand
4

Luciferase Assay of AHNAK-miR-1251

Check if the same lab product or an alternative is used in the 5 most similar protocols
The predicted 3'-UTR sequence of AHNAK binding to miR-1251 and the mutated sequence were inserted into the pGL3 promoter vector (Genechem, Shanghai, China) named pGL3-AHNAK-WT and pGL3-AHNAK-MUT. For reporter assay, cells were planted into 24-well plates and transfected with 100ng of pGL3-AHNAK-WT and pGL3-AHNAK-MUT, 50nM miR-1251 mimics and negative control using Lipofectamine 2000. Renilla luciferase vector pRL-SV40 (5 ng) was transfected into cells as control. Based on the obtained ratio, the activation degree of target reporter genes in different sample was compared.
Zhou L., Zhi Z., Chen P., Du C., Wang B., Fang X., Tang W., & Li H. (2020). lncRNA-RMST Functioned as a SOX2 Transcription Co-regulator to Regulate miR-1251 in the Progression of Hirschsprung's Disease.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!