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Pooled CRISPR Screen Analysis

Pooled CRISPR Screen Analysis Online Inquiry

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 Genome-wide Assembly and Annotation

A large-scale genetic loss-of-function experimental framework engineered to seek the equivalent of a few needles in a haystack is CRISPR screening. CRISPR screening makes it easier for a cell type to explore key genes or genetic sequences that induce a particular function or phenotype. CRISPR is a way of making double-strand cuts at directly aimed sites in DNA, as it is used by many scientists. When genomic DNA in cells produces such cuts, the cells utilize their DNA repair processes to fix the cuts. The repair method is typically imprecise, resulting in variations that throw the targeted gene out.

The CRISPR screen is an essential molecular target technique of high-throughput screening. An effective CRISPR screen begins with a well-designed library of sgRNA that targets the genes or loci of preference. Based on sequence and then synthesis, sgRNA is developed, accompanied by vector cloning or transcribed into RNA for cell system transfection. Then run a screen with a positive or negative selection that is widely utilized. Regarding physical segregation of cells into two or more communities, PCR amplification and hugely parallel sequencing will be carried out. The sgRNA and targeted genes of interest will be recognized after statistical analysis.

The Bioinformatics Analysis That We Can Provide

CD Genomics has established a cost-effective, efficient, and high-throughput tactic for CRISPR Screen sequencing by integrating amplicon-based next-generation sequencing to implement the growing demands of the scientific community. In order to assist people to evaluate gene functions in a high-throughput manner, our CRISPR Screen sequencing system can offer you direct and specific data about sgRNA and targeted gene analysis and functional enrichment assessment.

For high-throughput screening, CRISPR screen innovation can be utilized, a huge proportion of gene mutant cells can be produced, and the mutant cell library can be screened under the activation of various external conditions. It is possible to further define the connection between phenotype and genotype through high-throughput sequencing and bioinformatics evaluation. In addition, CRISPR/Cas9 library-based high-throughput screening also overwhelms the challenges of low transfection effectiveness and the capacity to inhibit gene expression only at the mRNA level faced by RNA interference screening innovation.  In the period of precision medicine, it has huge scientific clinical relevance and extensive application space.

Advantages of Pooled CRISPR Screen Analysis

Up to 104 tests per run, substantial multiplexing flexibility, and high-throughput sequencing

Ultra-deep amplicon sequencing or collected areas with a coverage of more than 1000X

Without bias, cost-effective and extremely sensitive identification stages

No requirement for time-consuming and laborious cloning phases

CD Genomics Pooled CRISPR Screen Analysis Pipeline

CD Genomics Pooled CRISPR Screen Analysis Pipeline

Bioinformatics Analysis Content

Analysis/Pipeline Details
Raw Data QC Gives 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
Validation of CRISPR Libraries Offer the CRISPR libraries themselves with verification using high-throughput sequencing
Analysis of Target Loci For phylogenetic and barcoding assessment, findings can be employed

How It Works

CD Genomics is a professional bioinformatics service provider with years of experience in NGS and long read sequencing (PacBio SMRT and Oxford Nanopore platforms) data analysis, integrated analysis services, database construction and other bioinformatics solutions.

How It Works

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

  1. Park RJ, Wang T, Koundakjian D, et al. A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors. Nature genetics. 2017, 49(2).
  2. Chen S, Sanjana NE, Zheng K, et al. Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. Cell. 2015, 160(6).
  3. Parnas O, Jovanovic M, Eisenhaure TM, et al.  A genome-wide CRISPR screen in primary immune cells to dissect regulatory networks. Cell. 2015, 162(3).
* For Research Use Only. Not for use in diagnostic procedures.
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