CRISPR-Cas9 Services: Knock-In and Knock-Out Service
Quick Turnaround CRISPR-Cas9 Knock-Out and Knock-In Services 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 Knock-In or Knock-Out Project
Unlock precise genome editing with our comprehensive CRISPR-Cas9 services. Our CRISPR knock-in and knock-out service delivers end-to-end solutions for targeted gene modification or deletion, transforming your research capabilities with cutting-edge precision. This advanced service builds upon our foundational offerings, including CRISPR vector production, sgRNA cloning, and CRISPR cassette production and donor vector construction design, to create a complete genome editing workflow. Achieve reliable, reproducible results with our expert CRISPR team.
CRISPR-Cas9 services lead times can change, so be sure to check our current lead times.
CRISPR Knock-Out and CRISPR Knock-In Services
CRISPR knock-out aims to disrupt the function of a gene, often by introducing mutations that render the gene non-functional, while CRISPR knock-in aims to insert a specific genetic sequence into the genome to achieve a desired genetic change or addition.
Our CRISPR knock-out or knock-in services are a whole product solution for researchers to knock-out or knock-in a gene of interest. Our production deliverables contain a plasmid that expresses a specific gRNA and Cas9 protein, a negative, and a donor vector with Luciferase-CMV-GFP-T2A-Puro-PA cassette (unless otherwise noted) that allow drug selection of positive clones. Learn why a CRISPR-Cas9 service is preferrable to a DIY kit.
Not sure which service is right for your research? The table below outlines the key differences.
| CRISPR Knock-Out Service | |
|---|---|
| Objective | Knock-out productions disrupt or deactivate (“knock out”) the function of a specific gene by introducing small insertions or deletions (indels) into a target gene’s DNA sequence. These indels can introduce frame-shift mutations that often result in non-functional proteins or lead to premature stop codons. |
| Outcome | The gene’s function is disrupted and it may no longer produce a functional protein, effectively “turning off” the gene. |
| Applications | Knock-out genome editing is used to study gene function, model diseases, and explore the consequences of gene inactivation. It is also used as a therapeutic strategy for diseases caused by overactive or harmful genes. |
| CRISPR Knock-In Service | |
|---|---|
| Objective | Knock-in productions insert or add (“knock in”) a specific genetic sequence (usually a new or modified DNA segment) into a genome at a precise location, introducing a desired genetic change, such as adding a functional gene or correcting a mutated gene. |
| Outcome | A specific genetic sequence is integrated into the target gene or genomic location, leading to the expression of a modified or additional gene product. |
| Applications | Knock-in genome editing is used for various purposes, including introducing reporter genes, tagging proteins for tracking or visualization, correcting disease-causing mutations, or adding specific sequences for therapeutic purposes. |
Sample Submission Requirements for Knock-Out or Knock-In Services
To get started on your CRISPR-Cas9 Knock-Out or Knock-In production, our CRISPR team needs the following:
- Gene Target Information:
- Gene ID and Sequence
- Target species (e.g., human, mouse, etc.).
- Ensure access to the complete gene annotation, including exons, introns, and regulatory elements.
- Accession number or full gene sequence.
- Knock-Out Details
- Determine which exon or region to disrupt (e.g., start codon or functional domain).
- Consider off-target analysis to avoid unintended modifications.
- Knock-In Details
- Define the exact sequence to introduce (e.g., SNP, tag, or entire gene).
- Include homology arms (~500–1,000 bp on either side of the edit).
- Gene ID and Sequence
- CRISPR Design Information:
- Guide RNA (gRNA) Design
- Select target sites with minimal off-target potential using design tools (e.g., CRISPRscan, Benchling, or CHOPCHOP).
- Optimal gRNA features: GC content ~40–60%, minimal self-complementarity.
- Choose the Cas protein suitable for the application:
- We recommend SpCas9
- Guide RNA (gRNA) Design
- Cell or Organism Information
- Cell Line or Organism
- Species and cell line specifics (e.g., HEK293T, C2C12, hMSCs).
- Doubling time and transfection efficiency.
- Delivery Method
- Choose the most efficient delivery method based on the cell type:
- Lipid-based transfection (e.g., Lipofectamine)
- Electroporation/nucleofection
- Viral vectors (AAV, adenovirus, lentivirus, retrovirus).
- Choose the most efficient delivery method based on the cell type:
- Cell Line or Organism
- Knock-In Specifics
- Donor Template for Knock-In
- Single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA).
- Includes homology arms flanking the intended edit.
- Repair Pathway
- Homology-directed repair (HDR) or non-homologous end joining (NHEJ).
- Donor Template for Knock-In
- Validation Plan
- Genotyping Assays
- PCR amplification and sequencing (e.g., Sanger, NGS).
- Restriction enzyme digestion for indel detection.
- Phenotypic Validation
- Confirm the functional outcome of the edit using assays (e.g., Western blot for protein knockout, reporter assays for KI).
- Off-Target Analysis
- Assess potential off-target edits with in silico tools and/or experimental validation.
- Genotyping Assays
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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