CD Genomics, as one of the most competitive service providers in sequencing and data analysis, we use CRISPR technologies and advanced bioinformatics to help you explore the genetic information and decipher the essence of life. Our skills and experience in data analysis can meet customers' personalized data analysis needs.
Introduction of CRISPR sgRNA Analysis
For genome engineering, Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) systems, forming a dynamic immune system in bacteria, have been revised. CRISPR has quickly become the most common genome engineering technique due to its comparative straightforwardness and flexibility. Two elements are included in engineered CRISPR systems: a guide RNA (gRNA or sgRNA) and a CRISPR-associated endonuclease (Cas protein). The gRNA is a short synthetic RNA consisting of a Cas-binding scaffold sequence and a user-defined ~20 nucleotide spacer defining the genomic target to be altered. Thus, by merely altering the target sequence existing in the gRNA, one can shift the genomic target of the Cas protein.
The CRISPR-Cas9 gene targeting system, which requires two elements: a custom RNA guide (sgRNA, composed of target-specific crRNA sequence and tracrRNA) and a non-specific endonuclease related to CRISPR (Cas9), is a new method for genome engineering that allows scientists to modify genome parts by removing, adding, or altering a portion of the organism's DNA sequence. It is presently the simplest, most flexible, accurate, and efficient genetic manipulation technique that has many possible applications, such as improvement of medicine and crop seeds. To make maximum genome editing, CRISPR-Cas9 has also been adjusted and has pioneered the creation of targeted mutations.
The Bioinformatics Analysis That We Can Provide
In the experimental process of CRISPR, validation of edits is particularly important. As off-target mutation and not a modify in the target gene could occur, next-generation sequencing (NGS) is a convenient and high-throughput technique to verify for preferred mutations. For academia, medicine, and industry, CRISPR amplicon sequencing has become a standard verification technique. High-throughput CRISPR screening by performing PCR tests using primers flanking the target region is according to the concept of targeted amplicon sequencing. The most delicate technique for verifying mutations is targeted amplicon sequencing and the identification frequencies are as low as 0.01 percent.
CD Genomics has established an inexpensive, efficient, and high-throughput tactic to screen and validate CRISPR-Cas9-based mutations by utilizing next-generation amplicon-based sequencing to resolve the growing demands of the scientific community. Our next-generation CRISPR sequencing platform can provide you with direct and comprehensive details on the nature and variety of the mutations, such as knockout/knockin allele verification, cutting efficiency assessment of sgRNAs, homozygous and heterozygous detection, assessment of mutation bandwidths, and more.
Advantages of CRISPR sgRNA Analysis
The relative simplicity of its plasmid layout and construction is offered in addition to these other mutagenic methods such as ZFN and TALEN.
Conveniently programmable by shifting the sequence of guides.
With the monomeric Cas9 protein and any quantity of different sequence-specific gRNAs, multiplexed genetic modification with the CRISPR-Cas9 library can be effectively obtained.
CD Genomics Pooled CRISPR Screen Analysis Pipeline
Bioinformatics Analysis Content
|
Analysis/Pipeline |
Details |
| Raw Data QC |
Offers a list of SNPs and individuals filtered out from |
| Reference Alignment |
Classify similarity areas that may be the result of functional, structural, or evolutionary connections between the sequences |
| sgRNA Abundance Analysis |
Evaluate the ample supply of sgRNA |
| Differential analysis of abundances of sgRNAs |
Analysis of distinctions between sgRNA abundances |
How It Works
CD Genomics is a high-tech company specializing in multiomic data analysis. We provide services such as project design, data analysis, and database construction. With a focus on developing breakthrough products and services, we are a pioneer in the biotechnology industry, serving researchers and partners worldwide.
CD Genomics has over a decade of experience in sequencing and analysis. We can help you with your project from experimental data, through all analyses to intact report at competitive rates. If you have any questions about how we can help you, please get in touch. We look forward to working with you!
References
- Chen L, Wang S, Zhang YH, et al. Identify key sequence features to improve CRISPR sgRNA efficacy. IEEE Access. 2017, 5.
- Xu H, Xiao T, Chen CH, et al. Sequence determinants of improved CRISPR sgRNA design. Genome research. 2015, 25(8).
- Larson MH, Gilbert LA, Wang X, et al. CRISPR interference (CRISPRi) for sequence-specific control of gene expression. Nature protocols. 2013, 8(11).
