CRISPR Library Screening for Target Identification    

Creative Biogene's CRISPR Library Screening Service aims to utilize the CRISPR/Cas9 system to disrupt gene function at the genome level, assisting scientists in rapidly identifying target genes, understanding their functions, and discovering new drug targets.

Background

Overview of CRISPR Library Screening

In recent years, CRISPR technology has emerged as a transformative tool in the field of biology. CRISPR library screening is a high-throughput screening method based on the CRISPR/Cas9 gene editing system, designed to interfere with the genome using a large-scale sgRNA library to discover new drug targets associated with specific phenotypes or biological processes. This technology employs precisely designed sgRNA sequences introduced into target cells or organisms, facilitating screening under drug pressure or cell sorting conditions to identify genes closely associated with the target phenotype. It provides critical molecular targets and mechanistic insights for drug development and accelerates the refinement of gene function studies and disease models. At the same time, the application of CRISPR library screening technology faces challenges such as off-target effects, complex sgRNA design, standardization of screening conditions, limitations in library coverage, and complexity in data analysis.

Figure 1. CRISPR screens can be described as the biological model in which the screen is conducted and the perturbations introduced using CRISPR technology. (Picco G, et al., 2019)

Screening Service

Service Offerings

Creative Biogene's CRISPR Library Screening Service effectively addresses these challenges by integrating advanced bioinformatics analysis techniques and precise experimental design. Our platform enables precise design and validation of sgRNA libraries optimizes screening conditions and analyzes data comprehensively to ensure high reliability and repeatability of screening results, providing efficient and accurate target discovery and gene function research services for our customers.

CRISPR Library Screening Service Process

• Target Selection and Design: Initial selection of target genes or loci and design of specific sgRNA sequences to ensure efficient gene editing and minimal off-target effects.
• Oligonucleotide Pool Synthesis and Cloning: Synthesis of designed sgRNA sequences into oligonucleotide pools and cloning into appropriate vectors for subsequent screening experiments.
• Library Construction and Quality Control: Cloning of synthesized sgRNA sequences into vectors to construct complete sgRNA libraries, followed by quality control to ensure library complexity and uniformity.
• Lentivirus Packaging: Packaging of constructed sgRNA libraries into lentiviral vectors for efficient library delivery into target cells for screening.
• High-Throughput Screening: Conducting high-throughput CRISPR screening in target cells, including knockout, activation, or suppression screening to explore functional changes related to genes or pathways of interest.
• PCR Amplification and Deep Sequencing: PCR amplification of screened samples followed by deep sequencing analysis to assess the abundance and screening efficacy of each sgRNA.
• Result Validation and Analysis: Validation of screening results for accuracy, analysis of biological significance for each hit site, and exploration of potential applications to ensure the reliability and repeatability of screening outcomes.
• Downstream Research and Applications: Further gene function research, drug target discovery, or crop improvement based on screening results to support innovative developments in scientific research and applications.

Figure 2. CRISPR-SCREEN screening detailed process.

Feature

Technical Assurance

Our team employs sophisticated technical assurance measures to deliver high-quality and reliable CRISPR Library Screening Service, enhancing the efficiency and success of your research.

• Coverage and Uniformity of Library Plasmids: We utilize a dual-plasmid system for sgRNA library to ensure nearly 100% coverage. Our electroporation methods and highly efficient transformation cells ensure a consistent abundance of each sgRNA.
• Library Virus Infection Efficiency: We precisely control infection efficiency at 30% to avoid mixed effects of multiple sgRNA copy numbers within cells.
• Cell Experiment Coverage: We conduct library screening experiments with 500x cell coverage to ensure data reliability at each experimental point and accurate interpretation of results.

Advantages of Our Service

• Efficient Library Construction: Rapid construction of high-quality sgRNA libraries using state-of-the-art CRISPR and high-throughput synthesis technologies.
• Comprehensive Data Analysis: Detailed gene function and target information are provided through deep sequencing and bioinformatics analysis.
• Flexible Customization: Personalized library design and screening strategies tailored to specific customer requirements.
• Diverse Applications: Widely applicable in drug development, gene function research, disease mechanism exploration, and more.

Application

Application Directions

By establishing a whole-genome CRISPR library, researchers can extensively screen genes, enabling in-depth exploration of gene roles in disease occurrence, drug resistance, viral infections, and other aspects. This technology provides a powerful tool for revealing gene functions and discovering new therapeutic targets and accelerates advancements in life sciences research.

1. Drug Resistance Mechanism Studies: CRISPR library screening can elucidate molecular mechanisms of tumor cell resistance to specific drugs. Through whole-genome knockout libraries, researchers can identify key genes involved in drug resistance, such as TP53, SOCS6, nf1, and med12 discovered in Imatinib and Vemurafenib resistance studies.

2. Virus Infection Target Gene Screening: Used to identify host factors influencing virus infection, such as CD4, and CCR5 found in HIV and EBV infection studies, and tumor genes associated with EBV. These studies provide crucial information for understanding virus transmission and developing vaccine or treatment strategies.

3. Tumor Development and Therapeutic Target Discovery: CRISPR library screening helps researchers identify genes crucial for maintaining tumor cell growth and development. For instance, knocking out the ARID2 gene revealed its inhibitory role in liver cancer cell proliferation, providing a scientific basis for developing new anti-cancer therapeutic targets.

4. Drug Development and Immunotherapy: Used to identify new drug targets and immunotherapy targets. For example, discovering DCPS as a novel target for AML treatment and exploring the role of CMTM6 in PD-L1 regulation pave the way for developing more effective and personalized treatment strategies.

5. Non-coding Sequence Functional Studies: CRISPR library screening is also employed in studying non-coding sequence functions, such as uncovering new mechanisms in P53 enhancer function research, and deepening understanding of gene regulation and tumor development mechanisms.

Contact Us

How to Get Started?

1. Consultation and Design: Contact our experts to discuss your research needs and goals, and receive professional advice and design proposals.
2. Library Construction and Screening: Based on the design plan, construct the corresponding sgRNA libraries and conduct high-throughput screening.
3. Data Analysis and Validation: Perform deep sequencing and data analysis on screening results, providing comprehensive reports and experimental validation.

For more information on our CRISPR Library Screening Service or to request a quote, don't hesitate to get in touch with the expert team at Creative Biogene. We are dedicated to providing high-quality gene editing and exploration services to scientists worldwide, making your research more efficient and precise.