CRISPR-Cas9 Services: sgRNA Cloning Service
Quick Turnaround sgRNA Cloning for Research and Development
Molecular Cloning & Mutagenesis Projects Completed
Viral Vector Packaging Solutions Delivered
Plasmid DNA Preparations Successfully Produced
Let's Get Started On Your sgRNA Cloning
Our sgRNA cloning services are often part of our full service CRISPR knock-out and knock-in projects. But we do provide sgRNA cloning as a stand alone service.
You can begin production by submitting your sgRNA cloning requirements on our request a quote form.
If you are interested in our full service CRISPR knock-out or knock-in where we perform sgRNA cloning to match your research requirements, request a quote to get started.
Not sure where to start? No worries. Begin with our step-by-step process to determine the key components of your sgRNA cloning project.
Step 1: Determine Target Gene Details
- Gene Symbol: Precise gene identifier for exact targeting (e.g., BRCA1, TP53, KRAS).
- Gene Accession Number: Ensures genomic accuracy, typically from NCBI or Ensembl databases.
- Organism: Determines specific genome reference and cloning strategy. Common examples include human, mouse, zebrafish, drosophila.
- Chromosome Location: Provides additional precision for gene targeting. Format like “17q21.31” for specific chromosomal regions.
- Specific Genomic Coordinates (if known/applicable): Allows most precise sgRNA design. Typically in format chr (e.g., chr17:41196312-41277500).
Step 2: Decide CRISPR Experimental Design
- Editing Objective: Determines sgRNA design strategy and downstream experimental approach. Primary editing goal examples: gene knock-out, gene modification, precise mutation, pr gene insertion.
- Cas9 Variant Preference: Influences editing precision and potential off-target effects. Cas9 variant options: standard Cas9 or nickase Cas9.
- Vector Preference: Determines RNA production and delivery method. These can be custom vectors or produced from our vector collection (see below table).
| Vector | Description |
|---|---|
| pGPS-Cas-T2A-Puro | In vivo expression sgRNA and Cas9 |
| pGPS-Nickase-T2A-Puro | In vivo expression sgRNA and Nickase Cas9 |
| pGPS-T7-sgRNA | T7 promoter in vitro sgRNA production |
| pGPS-T7-Cas9 | T7 promoter in vitro Cas9 RNA production |
| pGPS-Cas | In vivo Cas9 expression |
Step 3: Choose sgRNA Design Specifications
Number of sgRNAs requested: Increases editing efficiency; typically 2-3 sgRNAs per target to ensure successful modification.
Preferred Target Region: Influences editing efficiency and potential functional outcomes. Common choices include: 5′ coding region, 3′ coding region, promoter region, exon, or intron.
Off-target Analysis Requirement: Ensures specificity and reduces unintended genomic modifications. Choices include standard, comprehensive, or none needed.
Optional Step 4: Provide Experimental Context
- Cell Line/Type: Determines potential challenges in transfection and editing; common types include HEK293, HeLa, primary cells.
- Delivery Method: Influences sgRNA and Cas9 introduction strategy. Common examples are transfection, viral transduction, and electroporation (transfection is most common for adherent cell lines).
- Downstream Validation Method: Confirms successful gene editing via RT-qPCR, sequencing, or western blot. Our CRISPR team can provide all of these services.
CRISPR Vector Sequencing Reports for IND Filing
Integrated Vector Sequencing Reports are a critical component of any Investigational New Drug application submitted to the FDA for gene therapy products. This analysis provides essential verification of the vector’s genetic integrity, identity, and stability. By documenting the complete nucleotide sequence of a therapeutic vector, identifying any potential sequence variants, and confirming genetic stability across manufacturing processes, this report addresses crucial regulatory requirements while demonstrating a commitment to product quality.
Our CRISPR team can provide Integrated Vector Sequencing Reports for CRISPR vectors, just ask when starting a project.
CRISPR-Cas9 Technology in Biotechnology Research and Development
CRISPR technology has a wide range of applications across various fields, such as precision medicine research, because of its versatility and precision in editing genes and DNA sequences.
Companies and organizations like Editas Medicine, CorriXR Therapeutics, and CRISPR Therapeutics are several companies who utilize CRISPR-Cas9 in their groundbreaking platforms.
Want to learn more about the latest in CRISPR genome-editing based research? Our colleagues at ScienceDirect and Genetic Engineering & Biotechnology News continuously collect and publish the latest information on CRISPR based research.
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