No knockout allele available for your gene of interest ?

Let our team create one for you.

Genome ProLab can help you generate a custom CRISPR-Cas9 knockout allele in Drosophila melanogaster. We tailor the strategy to your target gene, research objectives, and available genetic resources.

How Are Knockout Alleles Generated ?

CRISPR-Cas9 uses a guide RNA (gRNA) to direct the Cas9 nuclease to a selected genomic sequence. Cas9 creates a double-strand break, which the cell can repair through non-homologous end joining (NHEJ) without requiring a donor DNA template.

This repair process can introduce small insertions or deletions (indels). When these changes disrupt the coding sequence—for example, by creating a frameshift—they can lead to loss of gene function.

Alternatively, two gRNAs can be used to target sites flanking a selected region, allowing the recovery of a deletion that removes an essential part of the gene.

A Strategy Tailored to Your Gene

The most suitable approach depends on the gene’s structure, the desired mutation, and the resources already available.

Using existing gRNA stocks

When suitable gRNA-expressing fly stocks are available, they may be crossed with a compatible germline Cas9-expressing strain to generate heritable mutations. We assess whether the available guides target regions appropriate for your knockout objectives.

Designing custom gRNAs

When existing stocks are unavailable or unsuitable, custom gRNAs can be designed to target your gene. Depending on the project, the strategy may aim to generate disruptive indels or delete a selected genomic region.

Considering the genetic context

Our strategy also takes into account alternative transcripts, the required genetic background, and potential effects on neighboring genes. For essential genes, lethal or sterile alleles may need to be maintained using appropriate balancer stocks.

Screening and Molecular Characterization

Candidate alleles are screened and characterized by DNA sequencing to identify the resulting mutation and assess its predicted effect on gene function.

Because not every indel produces a functional knockout, allele selection is an important part of the process. Additional functional studies may be needed to establish whether an allele completely abolishes gene activity.

Tell Us About Your Target Gene

Send us your gene name or FlyBase ID, your research objectives, and any relevant fly stocks or gRNA resources you already have.

Our team will review your project and provide a proposed strategy and a customized quote.