AAV sgRNA Library
AAV sgRNA Library
An AAV sgRNA library is a pooled collection of single-guide RNAs (sgRNAs) packaged into adeno-associated virus (AAV) vectors, designed for high-throughput CRISPR-based genome editing screens. This powerful tool enables researchers to systematically investigate gene function, identify therapeutic targets, and dissect complex biological pathways in vitro and in vivo.
Leveraging the efficiency and tissue specificity of AAV delivery, AAV sgRNA libraries allow precise genetic perturbations in hard-to-transfect cells and relevant animal models, including non-dividing cells. These libraries can be customized to target whole genomes, gene families, pathways, or specific genomic regions, providing unparalleled versatility for functional genomics studies.
Compared to lentiviral sgRNA libraries, AAV sgRNA libraries offer unique advantages—stable expression in non-dividing cells, broad tropism, low toxicity, long-term in vivo expression, and compatibility with tissue-specific AAV capsids. This makes them especially suited for CNS, muscle, liver, immune system, and organoid-based functional genomics, as well as in vivo pooled screening in mice and non-human primates.
AAV sgRNA libraries are typically delivered as a pooled viral population, maintaining high diversity (10⁴–10⁶ sgRNAs or more). Each shRNA can be tracked using next-generation sequencing, enabling researchers to measure enrichment or depletion of specific guides after selection.
Applications
- Genome-wide and targeted loss-of-function screening
- Pathway discovery and modifier identification
- Synthetic lethality studies
- Target validation for drug development
- In vivo pooled CRISPR/RNAi screening
- Organoid-based functional genomics
End-to-End Library Services
AAVnerGene manages every step of the workflow:
Library design & sgRNA selection
High-coverage cloning and amplification
NGS diversity confirmation (optional)
AAV packaging at 10¹²–10¹⁴ vg scale
Quality control: titer, purity, full/empty ratio, and infectivity
Follow-up evolution or screening support
AAV sgRNA Library Design
At AAVnerGene, we offer two flexible and scalable approaches to design your AAV sgRNA library—whether you prefer to use an existing validated sgRNA collection or build a fully customized library from scratch. Our goal is to ensure optimal diversity, balanced representation, and high functional performance for your screening applications.
| Feature | From Existing sgRNA Collections | Fully Custom AAV sgRNA Library Design |
|---|---|---|
| Source of shRNAs | Pre-validated, published, or commercially available shRNA sets | Entirely custom-designed based on your target genes |
| Design Flexibility | Limited to sequences already available | Full control over target genes, number of shRNAs/gene, targeting region, GC content, off-target filtering |
| Complexity | Usually fixed based on the existing library (10³–10⁵) | Flexible—small focused libraries to large genome-wide screens (10²–10⁶+) |
| Turnaround Time | Fastest option; minimal design effort | Longer design and validation phase |
| Cost | Lower, due to no sequence design work | Higher, due to custom sequence design and synthesis |
| Performance Validation | Often includes prior functional data | Can incorporate computational scoring (on-target/off-target) and optimized RNAi rules |
| Applications | Standard pathway studies, well-known gene sets | Novel target discovery, unique gene panels, species-specific targets |
| Sequence Optimization | Limited | Fully optimized for structure, stability, and knockdown efficiency |
| Barcode Integration | Optional | Fully customizable barcoding strategy (1–10+ barcodes/shRNA) |
| Species Compatibility | Based on available collections | Designed for any species (human, mouse, rat, NHP, etc.) |
| Best For | Fast, cost-effective studies using known genes | High-precision, tailored functional genomics or novel discovery projects |
sgRNA oligo pool
Oligo Pools Pricing at AAVnerGene
Complexity | 20 – 120nt | 121 – 150nt | 151 – 200nt | 201 – 250nt | 251 – 300nt |
| 2-100 Oligos | $400.00 | $480.00 | $540.00 | $700.00 | $1,000.00 |
| 101-500 Oligos | $800.00 | $960.00 | $1,080.00 | $1,400.00 | $2,000.00 |
| 501-1,000 Oligos | $1,200.00 | $1,440.00 | $1,620.00 | $2,100.00 | $3,000.00 |
| 1,001-2,000 Oligos | $1,600.00 | $1,920.00 | $2,160.00 | $2,800.00 | $4,000.00 |
| 2,001-6,000 Oligos | $2,400.00 | $2,880.00 | $3,240.00 | $4,200.00 | $6,000.00 |
| 6,001-12,000 Oligos | $3,200.00 | $3,840.00 | $4,320.00 | $5,600.00 | $8,000.00 |
| 12,001-18,000 Oligos | $4,000.00 | $4,960.00 | $5,400.00 | $6,800.00 | $10,000.00 |
| 18,001-24,000 Oligos | $5,200.00 | $6,400.00 | $7,200.00 | $8,800.00 | $13,000.00 |
| 24,001-30,000 Oligos | $6,800.00 | $8,400.00 | $9,000.00 | $11,800.00 | $16,400.00 |
| 30,001-42,000 Oligos | $7,800.00 | $9,600.00 | $11,600.00 | $13,000.00 | $20,000.00 |
| 42,001-54,000 Oligos | $9,400.00 | $11,800.00 | $13,200.00 | $16,000.00 | $23,000.00 |
| 54,001-72,000 Oligos | $10,800.00 | $13,000.00 | $14,200.00 | $18,000.00 | $27,000.00 |
| 72,001-96,000 Oligos | $11,800.00 | $14,000.00 | $15,000.00 | $19,000.00 | $29,000.00 |
| 96,001-120,000 Oligos | $12,800.00 | $15,000.00 | $16,000.00 | $20,000.00 | $31,000.00 |
| 120,001-150,000 Oligos | $13,800.00 | $16,000.00 | $17,800.00 | $22,000.00 | $33,000.00 |
| 150,001-180,000 Oligos | $14,800.00 | $18,000.00 | $19,200.00 | $24,000.00 | $36,000.00 |
| 180,001-210,000 Oligos | $15,800.00 | $19,200.00 | $21,800.00 | $26,000.00 | $39,000.00 |
| 210,001-240,000 Oligos | $17,200.00 | $21,800.00 | $23,200.00 | $28,000.00 | $43,000.00 |
| 240,001-300,000 Oligos | $21,000.00 | $25,500.00 | $27,000.00 | $33,000.00 | $49,000.00 |
| 300,001-360,000 Oligos | $25,200.00 | $30,600.00 | $32,400.00 | $39,000.00 | $59,000.00 |
| 360,001-420,000 Oligos | $29,200.00 | $36,000.00 | $38,000.00 | $46,000.00 | $70,000.00 |
| 420,001-480,000 Oligos | $33,600.00 | $41,000.00 | $43,200.00 | $52,800.00 | $82,000.00 |
| 480,001-600,000 Oligos | $41,200.00 | $50,000.00 | $53,200.00 | $64,800.00 | $100,000.00 |
| 600,001-696,000 Oligos | $48,800.00 | $60,000.00 | $63,000.00 | $78,000.00 | $114,000.00 |
AAV sgRNA Library Construction
| Component | Options | Default Recommended |
|---|---|---|
| shRNA Promoter | U6, H1, tRNA, | U6 |
| Cas Promoter | MP135, MP84, CMV, EF1a, hSyn….. | MP135 |
| CRISPR System | SpCas9, SaCas9, Cas12, Cas13 | No |
| AAV Genome | ssAAV or scAAV | ssAAV |
| Production System | AAVone, AAVdual, AAVtri | AAVtri |
| Barcoding | 0–10+ barcodes per sgRNA | No |
AAVnerGene offers backbone for different CRISPR/Cas systems.
AAV sgRNA library construction can be done using a variety of methods, including restriction enzyme digestion, ligation-independent cloning, or Gibson assembly. Our pricing is dependent on quantity and transfection conditions. Pricing increases with lower copies per cell.
Library complexity | Price | Turnaround Time |
1.00E+07 | $12,800 | 2-3 weeks |
1.00E+08 | $21,800 | 2-3 weeks |
| 1.00E+09 | $68,000 | 3-4 weeks |
| Higher | Please contact us to discuss | |
Note: Complexity is calculated based on the number of clones obtained after transformation. We also provide NGS (Next-Generation Sequencing) services to verify library diversity, available for an additional fee.
AAV sgRNA Library Production
Packaging of AAV shRNA Library follows the same process as standard AAV packaging. Please contact us (customer@aavnergene.com) to request a customized quote.
AAV sgRNA Library Validation
Validation of the AAV sgRNA Library is conducted using next-generation sequencing (NGS) to quantitatively assess enhancer integrity, diversity, and accurate enhancer-barcode pairing.
AAV sgRNA Library Evolution
- Library Packaging & Preparation
- Package the pooled sgRNA library into desire AAV vectors using high-efficiency production systems
- Quantify and validate the library diversity via NGS.
- Initial Transduction
- Infect target cells or animals with the AAV-sgRNA library at a low multiplicity of infection (MOI) to ensure mostly single vector integration per cell.
- Allow sufficient time for gene editing and phenotype manifestation.
- Selection Pressure Application
- Apply selective conditions relevant to your study (e.g., drug treatment, environmental stress, or disease model).
- This step enriches for cells with advantageous or resistant genotypes.
- Sample Collection & DNA Extraction
- Collect surviving or phenotypically altered cells or tissues.
- Extract genomic DNA for sgRNA amplification.
- Next-Generation Sequencing (NGS)
- Amplify sgRNA regions and perform deep sequencing to quantify sgRNA abundance.
- Compare to initial library to identify enriched or depleted guides.
- Data Analysis & Hit Identification
- Analyze sgRNA frequency changes to pinpoint candidate genes influencing the phenotype.
- Library Repackaging for Next Round
- Optionally, reconstruct an enriched sgRNA pool from hits for a subsequent round of evolution.
- Package this refined pool into AAV for the next selection cycle.
- Iterative Cycles
- Repeat steps 2–7 for multiple rounds to progressively enrich the most effective sgRNAs.
- Each cycle increases the robustness and confidence of your screen.
AAV sgRNA Library Services at AAVnerGene
AAVnerGene offers comprehensive AAV sgRNA library services, covering the entire workflow from custom library design and high-quality AAV packaging to multiple rounds of library evolution and analysis.
Our Services Include:
Custom sgRNA Library Design: Genome-wide, pathway-specific, or focused libraries designed with optimal guide selection and barcode integration.
High-Quality AAV Packaging: Utilizing our proprietary AAVone® and AAVdual® systems to ensure high titer, purity, and consistent transduction.
Library Validation: Next-generation sequencing (NGS) to confirm diversity and representation of sgRNAs.
Iterative Library Evolution: Support for multi-round in vitro or in vivo selection to enrich functional sgRNAs under relevant biological conditions.
Data Analysis & Reporting: Bioinformatic support to identify enriched guides and interpret screening results.
Partner with AAVnerGene to harness the full potential of AAV-delivered CRISPR technologies and drive impactful gene discovery and therapeutic innovation.