Results: siRNA sequences designed for Gene A and Gene B with GenScript’s bioinformatic design platform achieved higher knock down efficiency (KD%) than those created with other vendors’ tools.
One stop service
Flexible solutions
Experience-enabled fast delivery
| Service | Length | Quantity | Type | Purification | Modification | Conjugation | QC * | Function Testing # |
|---|---|---|---|---|---|---|---|---|
| siRNA | ~21-23nt | 5 nmol to Kg level |
|
Rnase free HPLC, desalt, PAGE | Locked Nucleic Acid (LNA), thioate, methylation, 2-Ome etc. (~200 types) | Antibody, peptide, lipid etc. |
|
|
| ASO | ~30nt |
|
Final Deliverables
| QC Item | Test Method | Specification | Standard QC | Customized QC |
|---|---|---|---|---|
| Appearance | Visual Inspect | White to pale yellow powder | ||
| Identity by MW | Mass Spectrometry | Theoretical Average MW ± 0.1% | ||
| LC-MS | Liquid chromatography–mass spectrometry | Report result | ||
| LC MS/MS Sequencing | High-resolution Mass Spectrometry Sequencing | 100% sequence coverage | ||
| Purity by HPLC | High Performance Liquid Chromatography | double-strand siRNA ≥ 90% | ||
| Purity by CGE | Capillary Gel Electrophoresis | Report result | ||
| Purity by PAGE | Polyacrylamide Gel Electrophoresis | Report result | ||
| pH Measurement | Micro pH Electrode Method | Report result | ||
| TEA Salt residue detection | Gas chromatography-mass spectrometry Measurement | Report result | ||
| Solvent residue | Gas chromatography-mass spectrometry Measurement | Report result | ||
| Endotoxin | Chromogenic Method | Report result | ||
| Bacterial Testing | Direct Inoculation /membrane filter method | Report result |
Please find more customized QC information in Oligo Quality Control Service>>.
# Available cell line models for function testing >>
| Disease State or Tissue Type | Cell Line |
|---|---|
| Bladder cancer | RT-4 |
| Cervical cancer | Hela |
| Fibroblast cancer | SJCRH30 |
| Leukemia | THP1, Jurkat, SUP-T1 |
| Liver cancer | HepG2, Huh-7, Hep3B |
| Lung cancer | A549, H1975 |
| Neuroblastoma | SH-SY5Y, SK-N-BE(2) |
| Ovarian cancer | RMG-1 |
| Skin cancer | A375, A431 |
| Non-cancerous | HEK293T, HUVECS |
| Mouse | L929S, CHO, Raw264 |
| Primary cells | PBMC, Primary Cynomolgus Monkey Hepatocytes |
GenScript also support customized cell mode
Results: siRNA sequences designed for Gene A and Gene B with GenScript’s bioinformatic design platform achieved higher knock down efficiency (KD%) than those created with other vendors’ tools.
Results: Effective site-specific conjugation between siRNA and antibody (ARC, Antibody RNA Conjugation) not only ensure knock down efficiency but also help make targeted delivery possible
Results: siRNA knock down efficiency (KD%) with different delivery systems
Many of GenScript's designed siRNAs achieved better knockdown efficiency when compared to a positive control siRNA created from a published sequence. Out of 100+ sequences, we selected 20 preliminary screening sequences for further optimization.
The 20 preliminary screening sequences were optimized through different modifications to improve knockdown efficiency. After optimization, the IC50 is reduced to 1/10 of the positive control’s value.
ELISA assay
Application: suitable for quantification of secreted proteins
Advantages: qualitative as well as quantitative (Microplate reader)
Immunofluorescence Staining
Application: suitable for quantification of membrane proteins
Advantages: qualitative as well as quantitative (Flow cytometry)
Western blot
Application: suitable for quantification of all kinds of proteins
Advantages: qualitative, sometimes semi-quantitative (Gel electrophoresis)
RNAi utilizes short oligo sequences to target and modulate genetic material to inhibit or enhance the production of specific proteins involved in disease processes, enabling highly targeted therapy.
siRNA (small interfering RNA)
Mechanism: siRNA forms gene silencing complex RISC with proteins like AGO2, specifically cleaving and degrading target mRNA.
Characteristics:
ASO (Antisense oligonucleotides)
Mechanism: ASO binds to mRNA forming DNA/RNA double-stranded structure, RNAse H binds to it and degrades target mRNA
Characteristics:
| Characteristics | Small molecule drug | Antibody drug | RNAi drug | Gene therapy drug | |
|---|---|---|---|---|---|
| Properties | Agent | Chemical molecules | Protein | RNA | DNA/gene editing tools |
| Molecular weight | Low (<900 da) | High (~150 kda) | Medium (13-16 kda) | Medium | |
| Target number | ++ | + | ++++ | +++ | |
| Target type | Protein | Secreted and membrane protein | mRNA | DNA | |
| Selectivity | ++ | +++ | ++++ | +++ | |
| Half-life | + (~Hours-days) | ++ (~Days-week) | ++++ (~Months) | ++++(~Months) | |
| Immunogenicity | Low | High | Low | Low | |
| Gene insertion risk | No | No | No | Yes | |
| Research & development | Stability | Stable | Unstable | Unstable | Unstable |
| Delivery | Easy | Difficult | Difficult | Difficult | |
| Research | ~Months | ~Months | ~Weeks | ~Weeks | |
| Lead optimization | Slow | Slow | Rapid | Slow | |
| Manufacture | Easy | Difficult | Easy | Medium difficult | |
siRNA / ASO advantages:
