Drosophila CRISPR/Cas9 Genome Editing
The clustered regularly interspaced short palindromic repeat (CRISPR) system was found as a bacterial immune system against invading viruses and plasmids. The class II relying on a single Cas protein with multiple domains, CRISPR/Cas9 system has been confirmed over the past decade the most widely studied and efficient tool to selectively mutagenize specific regions of the genome, and allow sophisticated and detailed mechanistic studies to be performed in a variety of organisms including Drosophila.
CD BioSciences as one of the transgenic Drosophila strain designers, we use leading edge technology to facilitate your exploration in life science. Our unique platform can meet customers' personalized research needs. Our services range from oligonucleotides customization to CRISPR editing, as well as off-target effect analysis. Our professional Ph.D. teams monitor every step of the operations guaranteeing your entire projects.
How does CRISPR/Cas9 work as a gene editing tool?
In Drosophila, the CRISPR/Cas9 system has been used to interrupt, delete, and replace genes. Cas9-induced double-strand breaks (DSBs) trigger DNA repair through two cellular pathways, non-homologous end joining (NHEJ) introduces small insertions and deletions (indels), and homologous directed repair (HDR) mediates precise repair, both of which can be used in genome editing.
Compared with ZFN and TALEN, the CRISPR/Cas9 system has distinct advantages in Drosophila gene editing: (1) more potential PAM sites can be found in the genome; (2) we provide germline expression of Cas9 and sgRNA Drosophila mutant lines, which greatly improves efficiency; (3) gene editing in Drosophila using the CRISPR/Cas9 system has high success rate. The CRISPR/Cas9 system has been effectively used to achieve knockout of loss-of-function genes with mutagenesis rates as high as 75-99%.
Fig.1 CRISPR/Cas9 Genome Editing Strategic Planning Flowchart (Gratz SJ et al, 2015)
Our Drosophila CRISPR/Cas9 genome editing service include
- CRISPR Oligonucleotides Design and Synthesis
- Drosophila CRISPR Knock-out Service
- Drosophila CRISPR Knock-in Service
- Drosophila CRISPR-KO-attP-KI Service
- Drosophila CRISPRi/CRISPRa Service
- CRISPR/Cas9 Off-target Effect Analysis
High-quality customed CRIPSR oligonucleotides services including sgRNA, ssDNA and dsDNA are produced on our advanced platform.
We offer one-stop CRISPR/Cas9 Gene Knock-out (KO) Drosophila services, either conventional or conditional, allowing you create own flies transformants.
CRISPR/Cas9 uses a new model of DNA-RNA recognition for gene editing, which greatly reduces the difficulty of design and the threshold of gene knock-in. We provide nearly any DNA fragments insertion services, also knock-in large gene (>10 kb) in high efficiency.
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.
CRISPRi/CRISPRa are flexible tools researching gene function and metabolism network, especially in vivo that effectively prevents the cytotoxicity. Loss-of-function (LOF) and gain-of-function (GOF) strategies are complementary. Conditional genome-wide activation or interference of target genes transcription can be achieved through our Drosophila CRISPRi/CRISPRa platform.
Off-target effects remain a major concern for CRISPR/Cas9-mediated genome editing. However, off-target analysis of the CRISPR/Cas9 system in Drosophila is very challenging. We offer multiple sequencing platforms and analysis algorithms, including PCR-based and next-generation sequencing technologies. Different algorithms are suitable for different situations.
Workflow of Drosophila CRISPR/Cas9 genome editing
We offer a CRISPR Drosophila general service includes design, cloning, microinjection, screening, validation and population building to bring you genetically edited Drosophila. Just tell us the name of the gene and how you want to edit it. We propose the best strategies and complete the rest of the steps for you.
- STRATEGY - Consult our genome engineering design experts to establish an exact custom CRISPR/Cas9 gene editing strategy.
- sgRNA CONSTRUCTS - Select sgRNA target sites and synthesize sgRNA. If you want, you can provide the target sequences and vector and we will construct the plasmid for you. Or we offer one-stop sgRNA design and exclusive target sites services for your research needs. The efficiency is tested in S2 or Kc cell lines before making the gene editing Drosophila you want (optional).
- DONOR DNA CONSTRUCTS- Design and synthesize donor DNA (if needed).
- MICROINJECTION - Plasmid microinjection by our skilled injection specialist (always >150 embryos to fulfill the guarantee).
- VALIDATION - Screening and identification (via PCR and sequencing).
- DELIVERY - Deliver the final Drosophila model and validation report, which describes the details of the genome editing and shows evidence of the required editing.
Drosophila in vivo CRISPR/Cas9 genome editing platform has a broad usage in fundamental research and drug management. Click here if you want to learn more.
- Genetics and evolution research.
- Metabolism research.
- Development and cell biology research.
- Neuroscience research.
- Drug discovery.
- Insect management research.
In our Drosophila Center, the Cas9 protein of these transgenic Drosophila strains can be specifically expressed in germ cells according to your needs. Cas9 transgenic lines with different chromosomes are available, so fertile progeny can be obtained by using Cas9 transgenic lines with different chromosomes, and Cas9 can be easily removed by hybridization.
CD BioSciences is a specialist provider of gene editing solutions for in vivo and in vitro disease Drosophila model construction, target gene knock-out/knock-in/CRISPRi/CRISPRa and other research needs. With years of expertise, our scientists provide clients with high-quality and powerful Drosophila transgenic strategy to study disease and development mechanisms, gene function, drug screening, and treatment strategy, etc. If you have any questions or requirements, please feel free to contact us.
- Gratz SJ, et al. (2015). CRISPR-Cas9 Genome Editing in Drosophila. Curr Protoc Mol Biol. 111: 31.2.1-31.2.20.
For research use only. Not intended for any clinical use.