| スケール | 応用 | 機能力価保証 ⓘ | ボリューム | TAT | デフォルトQC | デフォルトQC |
|---|---|---|---|---|---|---|
| Purified | デフォルトQC | > 1E+8 TU/mL | 0.1 mL |
7営業日から（Duplex qPCR） new
5営業日から(P24 ELISA) |
|
|
| 1 mL | ||||||
| 5 mL | ||||||
| > 1E+9 TU/mL | 1mL | |||||
| Supernatant | > 2E+6 TU/mL | 0.2 mL | 5営業日から |
|
|
注意:
P24 ELISA: カプシドタンパク質（p24）を定量し、ウイルス粒子の総数（物理力価）を測定します。算出される機能的力価（TU/mL）は、p24力価式に基づく予測値であり、実際の機能的力価とは異なる場合があります。
Duplex qPCR: 標的細胞への形質導入が可能な感染性ウイルス粒子を直接測定・定量することで、正確な機能力価を提供します。
| # | Vector Name | Application | Promoter | Antibiotics Marker | Fluorescence Marker |
|---|---|---|---|---|---|
| 1 | pGS-LVV-EF1a-[ORF] | Gene overexpression, CAR-T | EF1a | / | / |
| 2 | pGS-LVV-EF1a-[ORF]-hPGK-PuroR | Gene overexpression | EF1a | PuroR | / |
| 3 | pGS-LVV-EF1a-[ORF]-hPGK-BSDR | Gene overexpression | EF1a | BSDR | / |
| 4 | pGS-LVV-EF1a-[ORF]-hPGK-BleoR | Gene overexpression | EF1a | BleoR | / |
| 5 | pGS-LVV-EF1a-[ORF]-hPGK-HygR | Gene overexpression | EF1a | HygR | / |
| 6 | pGS-LVV-EF1a-[ORF]-hPGK-NeoR | Gene overexpression | EF1a | NeoR | / |
| 7 | pGS-LVV-EF1a-[ORF]-hPGK-PuroR-P2A-EGFP | Gene overexpression | EF1a | PuroR | EGFP |
| 8 | pGS-LVV-EF1a-[ORF]-hPGK-BSDR-P2A-EGFP | Gene overexpression | EF1a | BSDR | EGFP |
| 9 | pGS-LVV-EF1a-[ORF]-hPGK-BleoR-P2A-EGFP | Gene overexpression | EF1a | BleoR | EGFP |
| 10 | pGS-LVV-EF1a-[ORF]-hPGK-HygR-P2A-EGFP | Gene overexpression | EF1a | HygR | EGFP |
| 11 | pGS-LVV-EF1a-[ORF]-hPGK-NeoR-P2A-EGFP | Gene overexpression | EF1a | NeoR | EGFP |
| 12 | pGS-LVV-EF1a-[ORF]-hPGK-PuroR-P2A-mCherry | Gene overexpression | EF1a | PuroR | mCherry |
| 13 | pGS-LVV-EF1a-[ORF]-hPGK-BSDR-P2A-mCherry | Gene overexpression | EF1a | BSDR | mCherry |
| 14 | pGS-LVV-EF1a-[ORF]-hPGK-BleoR-P2A-mCherry | Gene overexpression | EF1a | BleoR | mCherry |
| 15 | pGS-LVV-EF1a-[ORF]-hPGK-HygR-P2A-mCherry | Gene overexpression | EF1a | HygR | mCherry |
| 16 | pGS-LVV-EF1a-[ORF]-hPGK-NeoR-P2A-mCherry | Gene overexpression | EF1a | NeoR | mCherry |
| 17 | pGS-LVV-EF1a-[ORF]-hPGK-PuroR-P2A-BFP | Gene overexpression | EF1a | PuroR | BFP |
| 18 | pGS-LVV-EF1a-[ORF]-hPGK-BSDR-P2A-BFP | Gene overexpression | EF1a | BSDR | BFP |
| 19 | pGS-LVV-EF1a-[ORF]-hPGK-BleoR-P2A-BFP | Gene overexpression | EF1a | BleoR | BFP |
| 20 | pGS-LVV-EF1a-[ORF]-hPGK-HygR-P2A-BFP | Gene overexpression | EF1a | HygR | BFP |
| 21 | pGS-LVV-EF1a-[ORF]-hPGK-NeoR-P2A-BFP | Gene overexpression | EF1a | NeoR | BFP |
| 22 | pGS-LVV-CAG-[ORF]-hPGK-PuroR | Gene overexpression | CAG | PuroR | / |
| 23 | pGS-LVV-CAG-[ORF]-hPGK-BSDR | Gene overexpression | CAG | BSDR | / |
| 24 | pGS-LVV-CAG-[ORF]-hPGK-BleoR | Gene overexpression | CAG | BleoR | / |
| 25 | pGS-LVV-CAG-[ORF]-hPGK-HygR | Gene overexpression | CAG | HygR | / |
| 26 | pGS-LVV-CAG-[ORF]-hPGK-NeoR | Gene overexpression | CAG | NeoR | / |
| 27 | pGS-LVV-CAG-[ORF]-hPGK-PuroR-P2A-EGFP | Gene overexpression | CAG | PuroR | EGFP |
| 28 | pGS-LVV-CAG-[ORF]-hPGK-BSDR-P2A-EGFP | Gene overexpression | CAG | BSDR | EGFP |
| 29 | pGS-LVV-CAG-[ORF]-hPGK-BleoR-P2A-EGFP | Gene overexpression | CAG | BleoR | EGFP |
| 30 | pGS-LVV-CAG-[ORF]-hPGK-HygR-P2A-EGFP | Gene overexpression | CAG | HygR | EGFP |
| 31 | pGS-LVV-CAG-[ORF]-hPGK-NeoR-P2A-EGFP | Gene overexpression | CAG | NeoR | EGFP |
| 32 | pGS-LVV-CAG-[ORF]-hPGK-PuroR-P2A-mCherry | Gene overexpression | CAG | PuroR | mCherry |
| 33 | pGS-LVV-CAG-[ORF]-hPGK-BSDR-P2A-mCherry | Gene overexpression | CAG | BSDR | mCherry |
| 34 | pGS-LVV-CAG-[ORF]-hPGK-BleoR-P2A-mCherry | Gene overexpression | CAG | BleoR | mCherry |
| 35 | pGS-LVV-CAG-[ORF]-hPGK-HygR-P2A-mCherry | Gene overexpression | CAG | HygR | mCherry |
| 36 | pGS-LVV-CAG-[ORF]-hPGK-NeoR-P2A-mCherry | Gene overexpression | CAG | NeoR | mCherry |
| 37 | pGS-LVV-CAG-[ORF]-hPGK-PuroR-P2A-BFP | Gene overexpression | CAG | PuroR | BFP |
| 38 | pGS-LVV-CAG-[ORF]-hPGK-BSDR-P2A-BFP | Gene overexpression | CAG | BSDR | BFP |
| 39 | pGS-LVV-CAG-[ORF]-hPGK-BleoR-P2A-BFP | Gene overexpression | CAG | BleoR | BFP |
| 40 | pGS-LVV-CAG-[ORF]-hPGK-HygR-P2A-BFP | Gene overexpression | CAG | HygR | BFP |
| 41 | pGS-LVV-CAG-[ORF]-hPGK-NeoR-P2A-BFP | Gene overexpression | CAG | NeoR | BFP |
| 42 | pGS-LVV-CMV-[ORF]-hPGK-PuroR | Gene overexpression | CMV | PuroR | / |
| 43 | pGS-LVV-CMV-[ORF]-hPGK-BSDR | Gene overexpression | CMV | BSDR | / |
| 44 | pGS-LVV-CMV-[ORF]-hPGK-BleoR | Gene overexpression | CMV | BleoR | / |
| 45 | pGS-LVV-CMV-[ORF]-hPGK-HygR | Gene overexpression | CMV | HygR | / |
| 46 | pGS-LVV-CMV-[ORF]-hPGK-NeoR | Gene overexpression | CMV | NeoR | / |
| 47 | pGS-LVV-CMV-[ORF]-hPGK-PuroR-P2A-EGFP | Gene overexpression | CMV | PuroR | EGFP |
| 48 | pGS-LVV-CMV-[ORF]-hPGK-BSDR-P2A-EGFP | Gene overexpression | CMV | BSDR | EGFP |
| 49 | pGS-LVV-CMV-[ORF]-hPGK-BleoR-P2A-EGFP | Gene overexpression | CMV | BleoR | EGFP |
| 50 | pGS-LVV-CMV-[ORF]-hPGK-HygR-P2A-EGFP | Gene overexpression | CMV | HygR | EGFP |
| 51 | pGS-LVV-CMV-[ORF]-hPGK-NeoR-P2A-EGFP | Gene overexpression | CMV | NeoR | EGFP |
| 52 | pGS-LVV-CMV-[ORF]-hPGK-PuroR-P2A-mCherry | Gene overexpression | CMV | PuroR | mCherry |
| 53 | pGS-LVV-CMV-[ORF]-hPGK-BSDR-P2A-mCherry | Gene overexpression | CMV | BSDR | mCherry |
| 54 | pGS-LVV-CMV-[ORF]-hPGK-BleoR-P2A-mCherry | Gene overexpression | CMV | BleoR | mCherry |
| 55 | pGS-LVV-CMV-[ORF]-hPGK-HygR-P2A-mCherry | Gene overexpression | CMV | HygR | mCherry |
| 56 | pGS-LVV-CMV-[ORF]-hPGK-NeoR-P2A-mCherry | Gene overexpression | CMV | NeoR | mCherry |
| 57 | pGS-LVV-CMV-[ORF]-hPGK-PuroR-P2A-BFP | Gene overexpression | CMV | PuroR | BFP |
| 58 | pGS-LVV-CMV-[ORF]-hPGK-BSDR-P2A-BFP | Gene overexpression | CMV | BSDR | BFP |
| 59 | pGS-LVV-CMV-[ORF]-hPGK-BleoR-P2A-BFP | Gene overexpression | CMV | BleoR | BFP |
| 60 | pGS-LVV-CMV-[ORF]-hPGK-HygR-P2A-BFP | Gene overexpression | CMV | HygR | BFP |
| 61 | pGS-LVV-CMV-[ORF]-hPGK-NeoR-P2A-BFP | Gene overexpression | CMV | NeoR | BFP |
| 62 | pGS-LVV-TRE-[ORF]-EFS-rtTA-Advanced-hPGK-PuroR | Inducible expression, Tet-on | TRE | PuroR | / |
| 63 | pGS-LVV-TRE-[ORF]-EFS-rtTA-Advanced-hPGK-BSDR | Inducible expression, Tet-on | TRE | BSDR | / |
| 64 | pGS-LVV-TRE-[ORF]-EFS-rtTA-Advanced-hPGK-BleoR | Inducible expression, Tet-on | TRE | BleoR | / |
| 65 | pGS-LVV-TRE-[ORF]-EFS-rtTA-Advanced-hPGK-HygR | Inducible expression, Tet-on | TRE | HygR | / |
| 66 | pGS-LVV-TRE-[ORF]-EFS-rtTA-Advanced-hPGK-NeoR | Inducible expression, Tet-on | TRE | NeoR | / |
| 67 | pGS-LVV-TRE-[ORF]-EFS-tTA-Advanced-hPGK-PuroR | Inducible expression, Tet-off | TRE | PuroR | / |
| 68 | pGS-LVV-TRE-[ORF]-EFS-tTA-Advanced-hPGK-BSDR | Inducible expression, Tet-off | TRE | BSDR | / |
| 69 | pGS-LVV-TRE-[ORF]-EFS-tTA-Advanced-hPGK-BleoR | Inducible expression, Tet-off | TRE | BleoR | / |
| 70 | pGS-LVV-TRE-[ORF]-EFS-tTA-Advanced-hPGK-HygR | Inducible expression, Tet-off | TRE | HygR | / |
| 71 | pGS-LVV-TRE-[ORF]-EFS-tTA-Advanced-hPGK-NeoR | Inducible expression, Tet-off | TRE | NeoR | / |
| 72 | pGS-LVV-hU6-[scramble shRNA]-EF1a-PuroR | shRNA knockdown | hU6 | PuroR | / |
| 73 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BSDR | shRNA knockdown | hU6 | BSDR | / |
| 74 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BleoR | shRNA knockdown | hU6 | BleoR | / |
| 75 | pGS-LVV-hU6-[scramble shRNA]-EF1a-HygR | shRNA knockdown | hU6 | HygR | / |
| 76 | pGS-LVV-hU6-[scramble shRNA]-EF1a-NeoR | shRNA knockdown | hU6 | NeoR | / |
| 77 | pGS-LVV-hU6-[scramble shRNA]-EF1a-PuroR-P2A-EGFP | shRNA knockdown | hU6 | PuroR | EGFP |
| 78 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BSDR-P2A-EGFP | shRNA knockdown | hU6 | BSDR | EGFP |
| 79 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BleoR-P2A-EGFP | shRNA knockdown | hU6 | BleoR | EGFP |
| 80 | pGS-LVV-hU6-[scramble shRNA]-EF1a-HygR-P2A-EGFP | shRNA knockdown | hU6 | HygR | EGFP |
| 81 | pGS-LVV-hU6-[scramble shRNA]-EF1a-NeoR-P2A-EGFP | shRNA knockdown | hU6 | NeoR | EGFP |
| 82 | pGS-LVV-hU6-[scramble shRNA]-EF1a-PuroR-P2A-mCherry | shRNA knockdown | hU6 | PuroR | mCherry |
| 83 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BSDR-P2A-mCherry | shRNA knockdown | hU6 | BSDR | mCherry |
| 84 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BleoR-P2A-mCherry | shRNA knockdown | hU6 | BleoR | mCherry |
| 85 | pGS-LVV-hU6-[scramble shRNA]-EF1a-HygR-P2A-mCherry | shRNA knockdown | hU6 | HygR | mCherry |
| 86 | pGS-LVV-hU6-[scramble shRNA]-EF1a-NeoR-P2A-mCherry | shRNA knockdown | hU6 | NeoR | mCherry |
| 87 | pGS-LVV-hU6-[scramble shRNA]-EF1a-PuroR-P2A-BFP | shRNA knockdown | hU6 | PuroR | BFP |
| 88 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BSDR-P2A-BFP | shRNA knockdown | hU6 | BSDR | BFP |
| 89 | pGS-LVV-hU6-[scramble shRNA]-EF1a-BleoR-P2A-BFP | shRNA knockdown | hU6 | BleoR | BFP |
| 90 | pGS-LVV-hU6-[scramble shRNA]-EF1a-HygR-P2A-BFP | shRNA knockdown | hU6 | HygR | BFP |
| 91 | pGS-LVV-hU6-[scramble shRNA]-EF1a-NeoR-P2A-BFP | shRNA knockdown | hU6 | NeoR | BFP |
| 92 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-PuroR | CRISPR engineering, Knockout, CRISPRi | hU6 | PuroR | / |
| 93 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BSDR | CRISPR engineering, Knockout, CRISPRi | hU6 | BSDR | / |
| 94 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BleoR | CRISPR engineering, Knockout, CRISPRi | hU6 | BleoR | / |
| 95 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-HygR | CRISPR engineering, Knockout, CRISPRi | hU6 | HygR | / |
| 96 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-NeoR | CRISPR engineering, Knockout, CRISPRi | hU6 | NeoR | / |
| 97 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-PuroR-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | PuroR | EGFP |
| 98 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BSDR-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | BSDR | EGFP |
| 99 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BleoR-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | BleoR | EGFP |
| 100 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-HygR-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | HygR | EGFP |
| 101 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-NeoR-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | NeoR | EGFP |
| 102 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-PuroR-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | PuroR | mCherry |
| 103 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BSDR-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | BSDR | mCherry |
| 104 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BleoR-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | BleoR | mCherry |
| 105 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-HygR-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | HygR | mCherry |
| 106 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-NeoR-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | NeoR | mCherry |
| 107 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-PuroR-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | PuroR | BFP |
| 108 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BSDR-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | BSDR | BFP |
| 109 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-BleoR-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | BleoR | BFP |
| 110 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-HygR-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | HygR | BFP |
| 111 | pGS-LVV-hU6-[scramble sgRNA]-EF1a-NeoR-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | NeoR | BFP |
| 112 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-PuroR | CRISPR engineering, Knockout, CRISPRi | hU6 | PuroR | / |
| 113 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-BSDR | CRISPR engineering, Knockout, CRISPRi | hU6 | BSDR | / |
| 114 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-BleoR | CRISPR engineering, Knockout, CRISPRi | hU6 | BleoR | / |
| 115 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-HygR | CRISPR engineering, Knockout, CRISPRi | hU6 | HygR | / |
| 116 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-NeoR | CRISPR engineering, Knockout, CRISPRi | hU6 | NeoR | / |
| 117 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-EGFP | CRISPR engineering, Knockout, CRISPRi | hU6 | / | EGFP |
| 118 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-mCherry | CRISPR engineering, Knockout, CRISPRi | hU6 | / | mCherry |
| 119 | pGS-LVV-hU6-[scramble sgRNA]-EFS-SpCas9-NLS-P2A-BFP | CRISPR engineering, Knockout, CRISPRi | hU6 | / | BFP |
| 120 | pGS-LVV-EF1a-dCas9-KRAB-MeCP2-P2A-PuroR | CRISPR engineering, CRISPRi | EF1a | PuroR | / |
| 121 | pGS-LVV-EF1a-dCas9-KRAB-MeCP2-P2A-BSDR | CRISPR engineering, CRISPRi | EF1a | BSDR | / |
| 122 | pGS-LVV-hU6-[scramble sgRNA]-EFS-dCas9-KRAB-MeCP2-P2A-PuroR | CRISPR engineering, CRISPRi | hU6 | PuroR | / |
| 123 | pGS-LVV-hU6-[scramble sgRNA]-EFS-dCas9-KRAB-MeCP2-P2A-BSDR | CRISPR engineering, CRISPRi | hU6 | BSDR | / |
| 124 | pGS-LVV-EF1a-dCas9-VP64-T2A-PuroR | CRISPR engineering, CRISPRa | EF1a | PuroR | / |
| 125 | pGS-LVV-EF1a-MS2-P65-HSF1-T2A-BSDR | CRISPR engineering, CRISPRa | EF1a | BSDR | / |
| 126 | pGS-LVV-EF1a-[ORF]-P2A-EGFP | Gene overexpression, CAR-T | EF1a | / | EGFP |
| 127 | pGS-LVV-EF1a-[ORF]-T2A-mCherry | Gene overexpression, CAR-T | EF1a | / | mCherry |
| 128 | pGS-LVV-EF1a-Kozak-EGFP-miR-30E(shRNA)-hPGK-PuroR | shRNA knockdown | EF1a | PuroR | / |
| 129 | pGS-LVV-EF1a-[ORF]-miR-30E(shRNA)-hPGK-PuroR | shRNA knockdown | EF1a | PuroR | / |
| 130 | pGS-LVV-EF1a-[ORF]-hPGK-PuroR-P2A-NLS-mCherry | Gene overexpression | EF1a | PuroR | NLS-mCherry |
| 131 | pGS-LVV-hU6-[scramble sgRNA] MS2 scaffold-hPGK-BleoR | CRISPR engineering, CRISPRa | hU6 | BleoR | / |
| 132 | pGS-LVV-EF1a-Firefly Luciferase-hPGK-PuroR | Gene overexpression | EF1a | PuroR | / |
| Bright Field | Fluorescence Field | |
 |
 |
Positive control MOI = 2 (by qPCR titration) |
 |
 |
Sample 1 LTR Insert Length: >9 kb Duplex qPCR Titer: 2.52×10^8 TU/mL MOI = 1.5 (by qPCR titration) |
 |
 |
Sample 2 - 1μL LTR Insert Length: >9 kb Duplex qPCR Titer: 2.06×10^8 TU/mL MOI = 1.5 (by qPCR titration) |
Challenge: Primary cells are difficult to transduce due to their non-dividing nature. And the downstream application (CAR-T therapy) required high transduction efficiency at a low MOI.
Solution: Built on our proprietary lentiviral packaging system and pGS-LVV vector, we optimized the transfection workflow with production boosters to deliver lentiviral vector with high functional titer.
Lentiviral Vector: pGS-LVV-EF1a-CD19CAR-T2A-EGFP
Result: Delivered lentiviral vector with a functional titer of 7.07×10⁹ TU/mL (duplex qPCR). The lentiviral vector successfully mediated anti-CD19 CAR expression in primary human PBMCs. Even at a low MOI of 1, transduction efficiency exceeded 70%.
| Human PBMC | Positive Cells (%) EGFP+/CD19CAR+ |
|---|---|
| Non-transduced | 0.0% |
| MOI=1 | 72.6% |
| MOI=2 | 69.0% |
| MOI=5 | 73.0% |
| MOI=10 | 71.5% |
Challenge: The oversized insert (>10 kb) compromises lentiviral packaging efficiency, resulting in low viral titer and poor downstream performance.
Solution: Utilizing our proprietary lentivirus packaging system, we optimized each step of virus packaging specialized for large
insert
(>6.5 kb) to achieve high-titer delivery.
Result:
| Bright Field | Fluorescence Field | |
 |
 |
Positive Control 5’ LTR to 3’ LTR: 6,570 bp MOI = 2 (by qPCR titration) Time: 48h after transduction |
 |
 |
Sample 1 5’ LTR to 3’ LTR: 11,288 bp Duplex qPCR Titer: 1.03×10^8 TU/mL MOI = 2 (by qPCR titration) Time: 48h after transduction |
Challenge: The requirement for co-delivering Cas9 and gRNA resulted in a large insert size, compromising lentiviral packaging efficiency. Additionally, the inherent resistance of macrophages to transduction, driven by their robust antiviral mechanisms, required lentivirus with high functional titers.
Solution: Utilizing our proprietary lentivirus packaging system, we optimized each step of virus packaging specialized for large insert(> 6.5kb) to achieve high-titer delivery.
Result: Delivered Cas9+gRNA lentivirus with a functional titer >1×10⁹ TU/mL, successfully establishing stable cell pools with >90% gene knockout efficiency.
Validation of Knockout Efficiency by Western Blot
Background: Multi-transmembrane proteins are challenging to express on the cell surface due to complex folding and membrane integration.
Strategy: Using lentiviral packaging of pGS-LVV-CAG-T-hPGK-PuroR, HEK293T cells were efficiently transduced at MOI 5 and 10.
Results: This approach achieved 100% positive expression of the T (12-transmembrane) protein, with levels exceeding 100-fold above baseline without enrichment, demonstrating that viral delivery enables high-efficiency transduction and robust expression of difficult multi-transmembrane targets.
