Gene function analysis, target discovery and validation, assay development, and compound screening often need cell-based assays. Stable cell lines that are engineered to express a gene of interest via transgene integrations into the host genome provide an efficient approach to conduct such analysis. However, because establishment of such cell lines are tedious in procedures, time-consuming in development process, and expensive in terms of labor, reagents, and materials, as well as post-transfection validation of individual clones, it is always debatable whether is worthy of developing such stable cell lines in individual laboratories.  

Signosis now provides a series of gene expression- or luciferase reporter-ready stable cell lines in order to facilitate the studies.

 

Luciferase Reporter Stable Cell Lines

Gene function analysis, target discovery and validation, assay development, and compound screening often need cell-based assays.  Stable cell lines that are engineered to express a gene of interest via transgene integrations into the host genome provide an efficient approach to conduct such analysis.  At Signosis, we help researchers save time and labor in generating and validating stable cell lines so scientists can spend more effort on solving the big questions.  We offer a wide selection of validated luciferase reporter cell lines that measure transcription factor activity as a read-out for various signaling pathways.

Benefits:

Routine mycoplasma testing - All cell lines tested negative for mycoplasma. High sensitivity and responsiveness - Each cell line is validated to induce a strong reporter signal in response to stimuli. Consistent - TF reporter construct is stably integrated into the genome to avoid experimental/cell-to-cell variations. Time-saving - Cell line can be used for experiments right away to study different signaling pathways. Firefly Luciferase Substrate -  Click here to learn more about Signosis luciferase substrate.

EGFR Stably Expressing Cell Lines

EGFR mutations occur in many types of cancers including non-small cell lung cancer. Common mutations found in these patients are deletions in exon 19 (Del19) and L858R substitution in exon 21.  Patients with these mutations are sensitive to first (gefitinib or erlotinib) and second (afatinib) generation EGFR tyrosine kinase inhibitors (TKIs), whereas patients with wild type EGFR are not.  However, eventually, all patients become resistant to first and/or second-generation EGFR TKIs, and acquired T790M secondary mutation in exon 20 accounts for more than 50% of the acquired resistance.  Cells expressing EGFR with either L858R+ T790M or Del19+T790M double mutations are resistant to induced apoptosis in the presence of first or second-generation EGFR TKIs. 

Eight cell lines stably expressing HA-tagged EGFR have been established in the IL3-dependent Ba/F3 cell lines —WT EGFR, L858R EGFR mutant, L858R/T790M EGFR double mutant, Del19 (del747-752) EGFR mutant, Del19/T790M EGFR mutant, three ins20 (D770_N771insSVD, A763_Y764insFQEA, A767_dupASV) EGFR mutants —and Ba/F3 cell line expressing empty vector.  Also, two cell lines stably expressing HA-tagged EGFR have been established in HCC827 cell lines —Exon19del EGFR mutant, Exon19del/T790M EGFR double mutant—and HCC827 cell line expressing empty vector.

Due to their proliferation upon EGFR activation, sensitivity to TKIs, or resistance to induced apoptosis, these cell lines can be used to study the molecular mechanism underlying susceptibility of tumors to the drugs as well as screening and validating new TKIs.

FLT3 Stably Expressing Cell Lines

The FMS-like tyrosine kinase 3 (FLT3) also known as cluster of differentiation antigen 135 (CD135) or fetal liver kinase-2 (Flk2) is a class III receptor tyrosine kinase (RTK) that plays a key role in controlling survival, proliferation, and differentiation of hematopoietic cells and it widely expresses in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Mutations in FLT3 (both internal tandem duplications (ITD) and point mutations) are among the most common somatic mutations in AML patients and are associated with poor prognosis and outcome in those patients.

HER2 (ERBB2) Stably Expressing Cell Lines

The proto-oncogene human epidermal growth factor receptor 2 (HER2) gene also known as ERBB2 is a member of the ERBB/HER RTK family which is comprised of EGFR (EGFR/HER1/ERBB1), HER3/ERBB3, and HER4/ERBB4. This gene encodes for a transmembrane glycoprotein receptor with an intracellular domain with tyrosine kinase activity which is responsible for the activation of intracellular signal transduction pathways controlling epithelial cell growth, differentiation, and angiogenesis. Aberrations of the HER2 gene which usually leads to increased receptor signaling have been observed in many different types of cancer.

 

pCMV-EGFR Vectors

Epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor that plays a critical role in regulating cell division and death.  Mutations in EGFR gene that lead to overexpression of the protein have been associated with a number of different cancers. The EGFR gene is present on chromosome 7p11.2 and has 28 coding exons in which exons 5–7 and 13–16 code for the ligand-binding domain and exons 18–24 code for the TK domain.  Downstream effects of EGFR activation include modulation of three major pathways of RAS-RAF-MAPK, PI3K/AKT, and JAK/STAT pathways which can lead to cell survival, proliferation, and migration of cancer cells.

 

 

We offer a collection of unique cloning and expression cDNA vectors contain WT and mutated EGFR gene. These mutations are clinically approved and are under further investigation by many researchers. Our high-quality vectors are fully sequenced and are ready to use. We also offer custom cloning services for your specific construct needs.

 

Highlight

• A panel of EGFR Mutant Vectors - The panel includes more than 12 ready-to-use EGFR mutants.
• DYKDDDDK-tag for easy detection - This tag is engineered to downstream of EGFR gene to facilitate Western blot
• Perfect for stable cell line establishment - The hygromycin-resistant gene is embedded in the vector for stable cell line establishment.
• Custom service available - We also offer the custom service with the same list price. If you have any mutants in your mind, please let us know.

 

 

pLenti-MSCV-EGFR-GFP Vectors

Epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor that plays a critical role in regulating cell division and death.  Mutations in EGFR gene that lead to overexpression of the protein have been associated with a number of different cancers. The EGFR gene is present on chromosome 7p11.2 and has 28 coding exons in which exons 5–7 and 13–16 code for the ligand-binding domain and exons 18–24 code for the TK domain.  Downstream effects of EGFR activation include modulation of three major pathways of RAS-RAF-MAPK, PI3K/AKT, and JAK/STAT pathways which can lead to cell survival, proliferation, and migration of cancer cells.

We offer a collection of unique cloning and expression cDNA vectors contain WT and mutated EGFR gene. These mutations are clinically approved and are under further investigation by many researchers. Our high-quality vectors are fully sequenced and are ready to use. We also offer custom cloning services for your specific construct needs.

• Infect both dividing and nondividing cells.
• Super bright CopGFP under EF-1 promoter as a selection marker.
• Contain HIV-1 LTRs and the lentiviral packaging signal (Ψ).
• Specific elements (WPRE, cPPT, RRE) included for improving transgene expression.
• DYKDDDDK-tag engineered to downstream of EGFR gene for Western blot.

pMSCV-EGFR-GFP Vectors

Epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor that plays a critical role in regulating cell division and death.  Mutations in EGFR gene that lead to overexpression of the protein have been associated with a number of different cancers. The EGFR gene is present on chromosome 7p11.2 and has 28 coding exons in which exons 5–7 and 13–16 code for the ligand-binding domain and exons 18–24 code for the TK domain.  Downstream effects of EGFR activation include modulation of three major pathways of RAS-RAF-MAPK, PI3K/AKT, and JAK/STAT pathways which can lead to cell survival, proliferation, and migration of cancer cells.

We offer a collection of unique cloning and expression cDNA vectors contain WT and mutated EGFR gene. These mutations are clinically approved and are under further investigation by many researchers. Our high-quality vectors are fully sequenced and are ready to use. We also offer custom cloning services for your specific construct needs.

 

• EGFR under MSCV promoter for high expression in hematopoietic or stem cells. 
• Super bright CopGFP under EF1 promoter as a selection marker.
• DYKDDDDK-tag engineered to downstream of EGFR gene for Western blot.

 

 

GAL4-UAS-luc Reporter Stable Cell Lines

The GAL4-UAS system is a powerful technique for studying gene expression and function. It has also been adapted to study nuclear receptor chemical-binding functions. Signosis GAL4-UAS-luc Reporter Stable Cell Lines is a reliable, rapid, and sensitive method for analyzing the expression of your target gene upon exposure to related ligands or other stimuli.

 

Benefits:

• Highly specific–the responsiveness of this stable cell line is driven by the specific ligand-binding domain with low/no cross-reactivity with other members of the nuclear receptor.
• Less toxic – The chimeric receptors display less toxic effects on the cells.
• Highly sensitive – More than 50 fold induction in response to the corresponding stimuli.
• Routine mycoplasma testing - All cell lines tested negative for mycoplasma.

 

NR LBD-Driven GAL4 Reporter Stable Cell Lines

Nuclear receptors (NRs) are a superfamily of transcription factors that regulate key aspects of development, metabolism, and disease. In general,  they activate the transcription of sets of genes in response to the binding of cognate ligands. The classic modular domain organization of NRs includes a variable N-terminal domain (NTD), a centrally located zinc-finger DNA-binding domain (DBD), and a C-terminal ligand-binding domain  (LBD).

The GAL4-UAS system is a powerful technique for studying gene expression and function. It has also been adapted to study nuclear receptor chemical-binding functions. Signosis NR LBD-driven GAL4 reporter stable cell lines is a reliable, rapid, and sensitive method for analyzing the intracellular status of NRs upon exposure to drug candidates or other stimuli. These functional assays allow for measurement of receptor activation or inhibition by compounds and can be used to determine compound potency and selectivity.

 

Benefits:

• Highly specific–the responsiveness of this stable cell line is driven by the specific ligand-binding domain with low/no cross-reactivity with other members of nuclear receptors.
• Less toxic – The chimeric receptors display less toxic effects on the cells.
• Highly sensitive – More than 50 fold induction in response to the corresponding stimuli.
• Routine mycoplasma testing - All cell lines tested negative for mycoplasma.

 

Parental Cell Lines

Signosis offers some of the parental (Host) cells used for generating its stable cell lines. These cell lines can be used as a matched control for their related stable cell lines. All cell lines tested negative for mycoplasma.

Bioluminescent Tumor Cell Lines

The gene encoding firefly luciferase is now the gene of choice for in vivo and in vitro reporting of transcriptional activity in eukaryotic cells. Reporter genes either fluorescent or bioluminescent, are a low-cost alternative for real-time analysis of gene expression in small animal models. In fluorescent approaches, an external source of light is required for the excitation of the protein. In contrast, bioluminescent reporter proteins can only produce light by using the appropriate substrates.

Bioluminescent tumor cell lines are critical to cancer investigation, especially imaging of tumors in xenografted animals.  They are used for assessing the dissemination of tumor cells in the body of xenografted mice as well as tumor growth or regression under treatment.  Although various types of reporter genes encoding fluorescent proteins have been reported, stable expression of exogenous luciferase in tumor cells combined with systemic injection of luciferin provides an excellent tool for external optical imaging.  Signosis has generated a number of tumor cell lines expressing luciferase genes, facilitating evaluation of novel therapeutic modalities. These cell lines can be used to monitor carcinogenesis and tumor progression during metastasis or compare highly invasive and non-invasive tumor cells. It would be also a great tool for studying the relationship between tumor cells and their specific microenvironments during metastasis. In addition, bioluminescent tumor cell lines can be used to develop an in vitro luciferase activity-based assay, as a positive control for luciferase activity, or to test if experimental conditions affect luciferase expression.

Benefits:
•    Consistent - Exogenous gene is stably integrated into the genome to avoid experimental/cell-to-cell variations.
•    Time saving - Cell line can be used for experiments right away to study different signaling pathways.

Stably Expressing Cell Lines