Convenient Luminescence Signal Monitors RNAi Effect
The psiCHECK™-1 and psiCHECK™-2 Vectors are designed to provide quantitative and rapid optimization of RNA interference (RNAi). The vectors enable monitoring of changes in expression of a target gene fused to a reporter gene. In both vectors Renilla luciferase is used as the primary reporter gene, and the gene of interest is cloned into a multiple cloning region located downstream of the Renilla translational stop codon. Initiation of the RNAi process by synthetic siRNAs toward a gene of interest results in cleavage and subsequent degradation of the fusion mRNA. Measuring decreases in Renilla activity provides a convenient way of monitoring the RNAi effect.
In comparison with other fusion approaches (e.g., GFP or flag-tags), the Renilla luciferase approach offers more convenient and rapid quantitation with higher sensitivity. The psiCHECK™-1 Vector is recommended for use in monitoring RNAi effects in live cells. The changes in Renilla luciferase activity are measured with the EnduRen™ Live Cell Substrate (Cat.# E6481), which allows continuous monitoring of intracellular Renilla luminescence. The psiCHECK™-2 Vector contains a second reporter gene, firefly luciferase, and is designed for endpoint lytic assays. Introduction of firefly luciferase in the psiCHECK™-2 Vector allows normalization of Renilla luciferase expression, achieving robust and reproducible results.
psiCHECK™ Vector Advantages for RNAi Optimization
- Convenient RNAi target evaluation.
- Quantitation is performed with a common luminometer; no need to purchase expensive equipmenton.
- Protocols allow for measurements in live cells or crude cell lysates.
- No labor-intensive, time-consuming assays or waiting for phenotypic changes.
- No need for transfection normalization when using the psiCHECK™-2 Vector.
- GenBank® Accession Number AY535007
- psiCHECK™-2 Vector sequence text file
Patents and Disclaimers
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If the researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide the researcher with a full refund.
Researchers may use this product for research use only, no commercial use is allowed. “Commercial use” means any and all uses of this product and derivatives by a party for money or other consideration and may include but is not limited to use in: (1) product manufacture; and (2) to provide a service, information or data; and/or resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researchers may: (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence, and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use or transfer of this product or derivatives is authorized without the prior express written consent of Promega. In addition, researchers must either: (1) use luminescent assay reagents purchased from Promega for all determinations of luminescence activity of this product and its derivatives; or (2) contact Promega to obtain a license for use of the product and its derivatives. Researchers may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. With respect to any uses outside this label license, including any diagnostic, therapeutic or prophylactic uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA. This label license relates to Promega patents and/or patent applications on improvements to the luciferase gene.
U.S. Pat. No. 8,008,006 and European Pat. No. 1341808.
Certain applications of this product may require licenses from others.