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Drosophila CRISPR-KO-attP-KI Service

CRISPR-Cas9 technology has revolutionized the field of Drosophila research, providing researchers with unprecedented precision and versatility in genetic manipulation. CD BioSciences utilizes CRISPR gene editing to modify Drosophila genetics precisely, offering CRISPR-KO-attP-KI service to assist researchers in Drosophila-related studies.

Overview of Drosophila CRISPR-KO-attP-KI

The CRISPR-KO-attP-KI service employs the powerful CRISPR-Cas9 gene editing technology to not only knockout target sequences but also strategically introduce attP landing sites. Subsequently, leveraging the attP-PhiC31 method, this enables the seamless insertion of modified or exogenous sequences. This innovative approach ensures a versatile and efficient workflow for creating complex genetic modifications in Drosophila.

Workflow of CRISPR-KO-attP-KI.

Fig.1 Workflow of CRISPR-KO-attP-KI.

Applications of Drosophila CRISPR-KO-attP-KI

The CRISPR-KO-attP-KI technique follows a two-step strategy involving knockout followed by knock-in. This innovative approach ensures enhanced flexibility in gene editing, resulting in substantial cost savings. This method excels in tackling intricate and challenging genetic modifications, including but not limited to:

  • Gene Knockout Phenotypic Rescue: Restore phenotypes by editing target genes while preserving essential functions.
  • Homologous Gene Replacement: Achieve seamless replacement of highly similar homologous genes.
  • Multi-locus Gene Mutagenesis: Construct mutants with alterations at multiple gene loci.
  • Insertion of Complex Sequences: Effortlessly insert multiple or complex sequences into the genome.
  • Conditional Knockout using FRT Sequences: Implement conditional gene knockout using the FRT system.

Our Services

Equipped with advanced genetic engineering techniques, CD BioSciences' services are tailored to researchers and scientists seeking to unravel the complexities of genetics using the highly versatile CRISPR technology.

Preparation of gRNA, cas9mRNA, and Donor Plasmid DNA
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Injection into Wild-Type Drosophila
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Cloning of Target Insert Sequence into attB Donor Vector
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Injection into attP-RFP Drosophila Strain and PhiC31-Mediated Integration
  • Design the suitable gRNA sequences for a specific target gene, generate the necessary Cas9 enzyme with cas9mRNA for precise genome editing, and construct a donor plasmid with homology arms and an attP-RFP cassette for targeted gene disruption. Finally, combine all these elements to create the injection mixture.
  • Inject the injection mixture into early-stage wild-type Drosophila embryos, which will develop into Drosophila with the precise knockout of the target gene, replaced by the attP-RFP cassette.
  • Clone the desired insert sequence, which may be a modified rescue sequence or an exogenous sequence, into an attB donor vector to create a donor DNA sample.
  • Inject the ready donor DNA sample into the pre-existing attP-RFP Drosophila strain, using the PhiC31 integrase system to facilitate the integration of the insert sequence between the attP sites. This process replaces the original genomic sequence with the preferred insert sequence, creating the final Drosophila strain.

Want to Learn More?

CD BioSciences are committed to delivering solutions that drive progress in Drosophila research. With our Drosophila CRISPR-KO-attP-KI service, we provide the tools to unravel the genetic mysteries of Drosophila and unlock a deeper understanding of their biology. Contact us to learn more about our service and discuss how we can assist you in your Drosophila research endeavors.

Reference

  • Yu, J. J. S., et al. One-step CRISPR-Cas9 protocol for the generation of plug & play conditional knockouts in Drosophila melanogaster. STAR Protocols. 2021, 2(2), 100560.

For research use only. Not intended for any clinical use.

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