AAV-sgRNA
AAVnerGene's AAV-sgRNA Services
Whether you’re working on basic gene editing research or developing therapeutic applications, AAVnerGene’s AAV-sgRNA Design and Selection Services offer the expertise and tools needed to drive your project forward. Contact us today to discuss your project and learn more about how we can support your gene editing needs.
Why Choose AAVnerGene for sgRNA Services?
Precision Design: Our sgRNA designs are based on cutting-edge algorithms and extensive validation, ensuring high specificity and efficiency for gene editing.
Comprehensive Solutions: From sgRNA design to AAV packaging and production, we provide an end-to-end solution tailored to your research needs.
Advanced Screening Tools: Our sgRNA selection kits and screening services allow for rapid identification of the most effective sgRNA sequences, saving you time and resources.
Expert Support: With years of experience in AAV vector production and gene editing technologies, our team provides expert guidance throughout your project, ensuring success at every stage.
Custom AAV-sgRNA Design Services
Precision-engineered sgRNA designs for optimal gene editing efficiency and specificity. We provide tailored single-guide RNA (sgRNA) design services for diverse CRISPR systems, ensuring high on-target activity and minimal off-target effects. Our designs are optimized for compatibility with AAV delivery, making them ideal for in vivo and therapeutic applications.
CRISPR-Cas9 Systems
We design high-performance sgRNAs optimized for SpCas9 and SaCas9, considering:
✔ Target site accessibility
✔ Off-target minimization (via advanced computational prediction)
✔ Editing efficiency (validated by empirical data when available)
CRISPR-Cas12 Systems
For compact and versatile Cas12-based editing, we offer sgRNAs compatible with:
Cas12a (Cpf1) – Efficient for staggered cuts and multiplexed editing
Cas12f (Ultra-compact CasΦ) – Ideal for AAV packaging and tight genomic spaces
CRISPR-Cas13 Systems
Specialized designs for RNA-targeting applications (e.g., transcript knockdown, base editing):
Cas13a, Cas13b, Cas13d – Optimized for high RNA-binding specificity and minimal collateral activity
Other Emerging Gene Editing Systems
Stay ahead with custom RNA-guided designs for novel CRISPR nucleases, RNA base editors, and epigenetic modifiers.
The Cas protein and sgRNA are combined in a single plasmid. Cas protein under a small but strong ubiquitous promoter. sgRNA is controlled by U6 promoter.
At AAVnerGene, we offer specialized AAV-sgRNA Selection Services to support your gene editing research, providing custom solutions for various CRISPR and other gene editing systems. With a focus on precision, efficiency, and flexibility, our services cater to a wide range of research applications, from basic research to therapeutic development.
AAV-sgRNA backbone plasmids
| pAAVtri-SpCa9-U6-sgRNA | Streptococcus pyogenes Cas9 (SpCas9) – Most widely used, recognizes NGG (canonical PAM). |
| pAAVtri-SaCa9-U6-sgRNA | Staphylococcus aureus Cas9 (SaCas9) – Smaller than SpCas9, good for AAV delivery (PAM: NNGRRT). |
| pAAVtri-NmCa9-U6-sgRNA | Neisseria meningitidis Cas9 (NmCas9) – Longer PAM (NNNNGATT), useful for targeting specific sequences |
| pAAVtri-Cas12a-U6-sgRNA | Cas12a (Cpf1, e.g., Acidaminococcus & Lachnospiraceae variants) – Cuts DNA with T-rich PAM (TTTV), generates sticky ends. |
| pAAVtri-Cas12b-U6-sgRNA | Cas12b (BvCas12b) – Thermostable, used in diagnostics (e.g., SHERLOCK). |
| pAAVtri-Cas12f-U6-sgRNA | Cas12f – Extremely compact (~400-700 aa), useful for viral delivery. |
| pAAVtri-Cas13a-U6-sgRNA | Cas13a (LwaCas13a) – Targets RNA, used for RNA knockdown (no DNA edits) and diagnostics (e.g., SHERLOCK) |
| pAAVtri-Cas13b-U6-sgRNA | Cas13b (PspCas13b) – Used in RNA editing (REPAIR, RESCUE) and transcriptome engineering. |
| pAAVtri-Cas13d-U6-sgRNA | Cas13d (RfxCas13d) – Compact, highly efficient for RNA knockdown in mammalian cells. |
AAV-sgRNA Selection Kits
The AAV-sgRNA selection kits include 4 sgRNAs, 1 control sgRNA, and a GOI-mCherry fusion reporter plasmid, providing a comprehensive toolkit for effective gene knockdown and real-time monitoring of transduction and expression. The sgRNAs are expressed by the U6 promoter, while the Cas proteins are controlled by a small but strong ubiquitous promoter, all within a single plasmid. Additionally, the GOI-mCherry fusion reporter plasmid is included to monitor gene knockdown efficiency. Customers only need to provide the GOI information or sequence to be silenced and select the gene editing systems. Co-transfection of the AAV-sgRNA plasmids and the GOI-mCherry reporter plasmid allows researchers to quickly assess the effectiveness of each sgRNA.
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AAV-sgRNA Selection Services
We offer end-to-end AAV-sgRNA screening services combined with our AAV-sgRNA Selection Kits to simplify the identification and optimization of high-performance sgRNAs.
| Service | Price | Cell Line | Method and Description | Additional Costs |
| Standard Selection | $2,000 | HEK 293T | Confocal Imaging: monitor mCherry Reporter expression | No |
| qPCR: Quantify mRNA knockdown efficiency | $50/Sample | |||
| Flow Cytometry: Quantify mCherry reporter expression | $200/Sample | |||
| NGS: Validate knock-out efficiency | $200/Sample | |||
| Custom Selection | Quote | Quote | Quote | Quote |
AAV-sgRNA screening workflow:
- AAV-sgRNA Selection Kits:
- Gene editing systems and Gene editing targets.
- Cell Line:
- Standard Selection: HEK293T, easy to transfection and widely used.
- Custom Selection: Easy transfection cell lines.
- Transfection: Co-transfect pAAVtri-sgRNA and GOI-mCherry plasmids at a 10:1 ratio using our PEIone transfection reagent.
- High transfection efficiency is essential for accurate identification of effective sgRNA candidates and reliable knockdown assessment.
- Validation Methods:
- Confocal Imaging: Monitor mCherry expression, Flow Cytometry is available upon request).
- qPCR: Quantify GOI mRNA knockdown efficiency.
- NGS: The targeted genomic region is initially amplified via PCR and then sequenced using NGS.
- Combine top-performing sgRNAs:
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- Combine selected sgRNAs into a single AAV vector for enhanced gene editing if necessary
- AAV Packaging: Seamless transition from screening to AAV production as all the plasmids are AAV backbone.
- Co-transfect AAV-shRNA plasmids with the pRCap plasmid (which carries the AAV rep and cap genes) and the mini-pHelper-1.0 plasmid (which supplies adenoviral E2A, E4orf6, and VA RNA functions) into HEK 293T cells.