CRISPR/Cas Gene Editing Vector Services

Alfa Cytology offers comprehensive CRISPR/Cas gene editing vector services designed to provide precise genome editing tools for your research projects. Leveraging an advanced CRISPR vector construction platform and optimized design strategies, our senior scientists are dedicated to developing highly efficient and specific gene editing vectors tailored to your specific research needs. We provide full-process support ranging from gene knockout/knock-in and transcriptional regulation to genome-wide screening, ensuring customized solutions at every critical step of your project.

Introduction to CRISPR/Cas Gene Editing Vector

The CRISPR/Cas system is currently the most powerful genome editing tool, utilizing a guide RNA (sgRNA) to direct Cas proteins to induce double-strand breaks (DSBs) at specific genomic loci, which are then repaired by endogenous non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways to achieve gene knockout, knock-in, or point mutation. The NHEJ pathway predominantly results in gene knockout via the introduction of frameshift mutations, whereas the HDR pathway enables precise knock-in or point mutation when a donor template is provided. Furthermore, by employing catalytically dead Cas9 (dCas9) fused to transcriptional activation or repression effector domains, CRISPR technology also enables transcriptional activation (CRISPRa) or interference (CRISPRi), offering reversible regulatory tools for gene function studies, particularly suited for essential gene functional analysis and the dissection of complex gene regulatory networks. In oncology research, CRISPR/Cas vectors are widely utilized in applications such as functional genetic screening, tumor driver gene validation, drug resistance model construction, and synthetic lethal target discovery. Depending on the research objectives, the CRISPR/Cas system can be employed to generate permanent gene knockout cell lines (via NHEJ-mediated frameshift mutations), precise gene knock-in models (via HDR-mediated site-specific integration), or reversible transcriptional regulation of endogenous genes through CRISPRi/a.

Fig. 1 Different workflows used in CRISPR screening and mutagenesis. (Chehelgerdi M, et al.; 2024)

Our Services

Our CRISPR/Cas gene editing vector services support a wide range of Cas9 variants, vector backbones, and delivery formats, accommodating various editing types including gene knockout, knock-in, point mutation, and transcriptional regulation.

sgRNA Design and Vector Construction

Based on your research objectives, we design and construct CRISPR vectors carrying sgRNA sequences. Our services include:

• sgRNA target design utilizing advanced algorithms (3–5 sgRNAs per gene).
• Off-target effect prediction and evaluation.
• sgRNA duplex annealing and vector ligation.
• A broad selection of Cas9 variants: SpCas9, SaCas9, Cas9n (nickase), and dCas9.
• Multiple vector backbones: conventional plasmids, lentiviral vectors (e.g., lentiCRISPR v2), and AAV vectors.
• Selection markers: Puromycin, Blasticidin, Neomycin, GFP, and mCherry.
• Full-length sequencing validation to ensure 100% accuracy of the inserted sequence.

Vector Functional Validation

• All CRISPR vectors undergo rigorous functional validation, including:
• Sequencing confirmation of sgRNA insert accuracy.
• Editing efficiency testing in standard cell lines (via T7EI assay or sequencing).
• Provision of a comprehensive editing efficiency evaluation report.

Available CRISPR Systems

We also offer gene-edited cell line construction services, including transfection/transduction, monoclonal screening, editing efficiency validation (T7EI/sequencing), and delivery of the final cell line.

Specifications

Applications of CRISPR/Cas Vectors in Cancer Research

Workflow of CRISPR/Cas Gene Editing Vector Services

Project Consultation and Strategy Design

We discuss your research objectives, gene information, cell type, and editing requirements to design the optimal CRISPR vector construction and editing strategy.

sgRNA Design and Vector Construction

sgRNA target design, off-target prediction, annealing, ligation, and transformation are performed, followed by full-length Sanger sequencing verification.

Editing Efficiency Validation

The CRISPR vector is transfected into a standard cell line, and editing efficiency is assessed by T7EI assay or sequencing.

Delivery and Downstream Application Support

CRISPR plasmids, glycerol stocks, and validation reports are delivered, with seamless transfer to downstream services such as gene-edited cell line construction and in vivo editing studies.

Whether you are conducting whole-genome screening, validating a tumor driver gene, or building a drug-resistance model, Alfa Cytology delivers CRISPR/Cas vectors engineered for efficiency and specificity. From sgRNA design to editing efficiency validation, our team provides the technical foundation your cancer research demands. Please don't hesitate to contact us to discuss how we can support your next editing project

For research use only.