Custom circRNA Expression Vectors
abm offers a unique Service where we can synthesize and clone your custom circRNA sequence into our over-expression vector system. circRNAs can be over-expressed in mammalian cells by using non-repetitive regions of reverse complementary sequences - these sequences function by bringing the exon-flanking portion of introns in close proximity, thus promoting back-splicing into circRNA. To get started on your custom circRNA vector, simply click the Inquire button below.
Related Products:
Service Details
Core Services
| Service | Unit | Cat. No. | Price |
|---|---|---|---|
| Custom circRNA Non-Viral Expression Vector | 1.0 µg | C341 | $45.00 |
| Custom circRNA AAV Expression Vector | 1.0 µg | C342 | $45.00 |
| Gene Synthesis | Per bp | C098 | from USD $0.18/bp |
Controls & Add on Services
| Product or Service | Unit | Cat. No. | Price |
|---|---|---|---|
| pNV-CMV-circ-GFP-Control | 1.0 μg | CIR001 | $250.00 |
| pAAV-CMV-circ-Control | 1.0 µg | G2002 | $250.00 |
| Bacterial Agar Stab | 1.0 | C315 | $40.00 |
| Plasmid Amplification Service | 10-20 µg | C309 | $75.00 |
| Plasmid Amplification Service | 80-100 µg | C310 | $150.00 |
Supporting Data
Our data speaks for itself:
A) Three circRNAs were tested for over-expression in HEK293T cells
circRNA 1, 2 and 3 have been documented to be expressed in many tissues; notably circRNA 1 is expressed in kidney tissues (and thus in HEK293T cells) whereas circRNA 2 and 3 are not.
circRNA 1, 2 and 3 have been documented to be expressed in many tissues; notably circRNA 1 is expressed in kidney tissues (and thus in HEK293T cells) whereas circRNA 2 and 3 are not.
| circRNA1 | circRNA2 | circRNA3 | |
| Expression Tissues | Kidney, Liver, Brain, Small Intestine, Stomach, Lung, Spleen, Thymus, Placenta, Colon, Bone Marrow | Brain, Liver, Placenta, Colon, Heart, Spinal Cord, Uterus, Prostate, Lymph node, Tonsil, Blood | Liver, Prostate, Colon, Brain, Bone Marrow, Testis, Breast, Lung |
| Associated Diseases | N/A | Breast cancer, clear cell renal cell carcinoma, epithelial ovarian cancer, glioblastoma, hepatocellular carcinoma | Bladder cancer, hepatocellular carcinoma, renal cell carcinoma, hepatocellular carcinoma |
| circRNA Type | 5’ UTR | Exonic | Exonic |
B) circRNAs were cloned into abm’s circRNA Expression Vector
The circRNA Expression Vector utilizes an optimized complementary sequence-mediated circRNA circularization strategy – these sequences work to bring the exon flanking portion of introns in close proximity thus promoting highly efficient back-splicing into circRNA in vivo.
The circRNA Expression Vector utilizes an optimized complementary sequence-mediated circRNA circularization strategy – these sequences work to bring the exon flanking portion of introns in close proximity thus promoting highly efficient back-splicing into circRNA in vivo.
C) circRNA Expression Vectors were transfected into HEK293T cells
The circRNA Expression Vectors contain an eGFP cassette allowing for convenient assessment of transfection efficiency.
abm products used: DNAfectin™ Plus Transfection Reagent (Cat. No. G2500)
The circRNA Expression Vectors contain an eGFP cassette allowing for convenient assessment of transfection efficiency.
abm products used: DNAfectin™ Plus Transfection Reagent (Cat. No. G2500)
D) Total RNA was extracted from transfected cells
Total RNA was analyzed for integrity and purity by running samples on an agarose gel.
abm products used:
Column-Pure RNA Miniprep Kit (Cat. No. D518)
SafeView™ Classic Nucleic Acid Stain (Cat. No. G108)
Total RNA was analyzed for integrity and purity by running samples on an agarose gel.
abm products used:
Column-Pure RNA Miniprep Kit (Cat. No. D518)
SafeView™ Classic Nucleic Acid Stain (Cat. No. G108)
E) Total RNA was subjected to RT-qPCR using divergent primers
The standard method to detect circRNA expression is by RT-qPCR using divergent primers. These primers are designed in opposing directionality, and thus will only create a qPCR amplicon if correct circularization has occurred.
abm products used:
OneScript® Hot cDNA Synthesis Kit (Cat. No. G594)
BlasTaq™ 2X qPCR MasterMix (Cat. No. G891)
The standard method to detect circRNA expression is by RT-qPCR using divergent primers. These primers are designed in opposing directionality, and thus will only create a qPCR amplicon if correct circularization has occurred.
abm products used:
OneScript® Hot cDNA Synthesis Kit (Cat. No. G594)
BlasTaq™ 2X qPCR MasterMix (Cat. No. G891)
circRNA1 was successfully over-expressed in HEK293T cells. Cells transfected with Empty Vector Control (red) or circRNA1 Expression Vector (green) were compared. Earlier CT values and amplification curves of circRNA1 indicated successful over-expression of circRNA1 in HEK293T cells. Melt curve analysis indicated the amplicon detected was the same circRNA species in both transfected cell groups, confirming endogenous expression of circRNA1.
circRNA2 was successfully ectopically expressed in HEK293T cells. Cells transfected with Empty Vector Control (red) or circRNA2 Expression Vector (green) were compared. Amplification curve of circRNA2 only (green) indicated correct circRNA expression and no endogenous expression of this species in HEK293T cells.
circRNA3 was successfully ectopically expressed in HEK293T cells. Cells transfected with Empty Vector Control (red) or circRNA3 Expression Vector (green) were compared. Amplification curve of circRNA3 only (green) indicated correct circRNA expression and no endogenous expression of this species in HEK293T cells.
