Clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) nuclease systems have rapidly become the most widely used plant and animal genome editing tools, enabling diversification in agricultural applications. In recent years, the programmable nature of the CRISPR-Cas system has enabled the construction of large libraries of knockout mutants for forward genetic screening. High-throughput screening using CRISPR Screen technology allows for the generation of large numbers of mutant cells and the screening of mutant cell libraries under different external environmental stimuli. The relationship between phenotype and genotype can be further determined by high-throughput sequencing and bioinformatics analysis. The main advantages of screening using CRISPR are its ease of identifying disease-causing genes and its relatively high specificity. In addition, the possibility of multiplexing allows for the simultaneous targeting of multiple genes, thus overcoming genetic redundancy and enabling saturation mutagenesis using smaller populations.

Fig. 1. Workflow and design parameters for CRISPR knockout screening in plants. (Anas et al., 2023)

Our CRISPR screening sequencing solution

To address the need for CRISPR/Cas9 in accelerating agricultural research and crop improvement, CD Genomics offers high-throughput CRISPR screening sequencing solutions tailored for agricultural applications by utilizing next-generation sequencing (NGS). Our services cover the entire workflow, from experimental design and gRNA library construction to high-throughput sequencing and data analysis.

Our CRISPR screening sequencing service enables genotyping by sequencing integrated gRNAs and/or target genes, providing you with direct and detailed information about sgRNA and target gene analysis as well as functional enrichment analyses, helping plant and animal researchers to study gene function in a high-throughput manner. We aim to help our customers deepen their understanding of plant gene function and facilitate engineering programs for crop traits.

Applications of CRISPR screening sequencing

Our CRISPR screening sequencing platform allows for the analysis of plant and animal genomes at a higher resolution, providing you with vital information for crop improvement, plant and animal breeding, and gene function analysis. Our CRISPR screen sequencing solutions are widely used:

• Obtaining genome sequences
• Identifying plant and animal lines with mutant phenotypes
• Identifying candidate genes through gRNA and target site analysis
• Identifying novel mutants with interesting phenotypes
• Target sequence diversification
• Studying gene function

CRISPR screening sequencing workflow

Fig. 2. CD Genomics' CRISPR screening sequencing workflow for agriculture.

Sample requirements and preparation

• Sample type: Cell or DNA sample after positive/negative selection
• DNA sample: ~1.5 μg (concentration ≥30 ng/μL; OD260/280=1.8~2.0)
• Cell sample: 5×10⁶ cells

Our strengths and features

• Our team of scientists has deep expertise in CRISPR screen sequencing and its application in agriculture.
• Our laboratories are equipped with state-of-the-art sequencing platforms and advanced molecular biology facilities to ensure the generation of high-quality, accurate sequencing data to unravel the complexity of plant genomes.
• We offer end-to-end CRISPR screen sequencing services to streamline the CRISPR screen sequencing workflow.
• We adhere to strict quality control measures throughout the process, from sample processing to data analysis.

CD Genomics' CRISPR screening sequencing solutions reliably and cost-effectively identify sgRNAs and target genes of interest and perform deep sequencing and data analysis. Our expertise and commitment to quality are well-positioned to support agricultural researchers in their pursuit of innovative solutions. By leveraging our comprehensive services and cutting-edge technologies, researchers can unravel the complexities of plant genomes and drive advances in sustainable agriculture, disease resistance, increased yields, and nutritional quality. If you are interested, please feel free to contact us.

Reference

1. Gaillochet, Christophe, Ward Develtere, and Thomas B. Jacobs. "CRISPR screens in plants: approaches, guidelines, and future prospects." The Plant Cell. 33.4 (2021): 794-813.