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Single-guide RNA (sgRNA) is an RNA sequence that guides the Cas9 protein to specific loci within the genome. With the rapid development of RNA-based technologies, sgRNA library services have become increasingly feasible. As a biotech expert, Creative Biogene specializes in providing high-quality, customized sgRNA library solutions. Equipped with cutting-edge facilities, mature technology platforms, and an experienced team, we design gene-specific sgRNA constructs, synthesize high-throughput oligonucleotides, and clone them into lentiviral vectors. Leveraging years of expertise in gene editing, we offer faster delivery timelines and a superior service experience.
Efficient CRISPR screening starts with the precise design of sgRNA libraries targeting genes or loci of interest. By synthesizing oligo pools that match sgRNA designs and cloning them into expression vectors or transcribing them for transfection, accurate genome editing can be achieved.
Well-designed sgRNA libraries are essential for genome and metabolic pathway editing. When coupled with high-throughput screening, these libraries become indispensable tools in drug development, gene function analysis, molecular diagnostics, disease therapeutics, and crop breeding. Forward genetic screening allows unbiased discovery of genetic elements related to target phenotypes, with CRISPR libraries being widely employed for large-scale functional genomic studies.
Creative Biogene provides comprehensive sgRNA library customization, covering target-specific sgRNA construct design, high-throughput oligonucleotide synthesis via chip technology, lentiviral vector cloning, high-titer viral packaging, thorough quality assessment, and sequencing validation, as well as expert experimental consultation. We offer end-to-end CRISPR library solutions that ensure accuracy and reliability at every stage—empowering successful gene function studies, drug target identification, and high-throughput screening experiments.
Oligo synthesis
Design and synthesize short DNA sequences encoding the gRNAs.
Elution
Purify the synthesized oligos to remove impurities or unbound reagents.
Ligation of gRNA library
Insert the oligos into a vector backbone to create a pooled gRNA library.
Lentivirus library construction
Package the gRNA library into lentiviral particles for delivery into cells.
Identification of functional genes
Sequence integrated gRNAs and map them to target genes to identify those responsible for the phenotype.
Analysis of the positive cell pools
Collect and analyze cells showing the phenotype of interest.
Cell screening based on phenotype
Infect target cells with the virus and screen for desired phenotypic changes (e.g., growth, resistance, morphology).
CRISPR libraries are built using high-throughput synthesis, forming plasmid pools of sgRNAs targeting multiple loci:
We employ cutting-edge software to generate sgRNA sequences with minimal off-target effects and maximum efficiency, using multiple algorithms to ensure specificity and potency.
GC Content
Maintained at 40–80% to ensure sequence stability and high cutting efficiency.
Sequence Length
Controlled between 17–24 nt based on the chosen Cas variant to balance efficiency and specificity.
Off-Target Prediction
Bioinformatics screening identifies mismatches to optimize design.
Redundancy
Multiple sgRNAs per gene ensure successful knockout.
Quality Metrics
Standardization
Creative Biogene fully recognizes the critical role of downstream functional screening in translating gene perturbations into biological insights. As an extension of our offerings, we provide comprehensive technical consultation and optional integrative support to assist clients in deploying sgRNA libraries within high-throughput screening workflows. These applications typically begin with the successful transduction of our high-titer lentiviral sgRNA libraries into target cells, followed by phenotypic selection or enrichment, PCR-based sgRNA amplification, and next-generation sequencing (NGS) to identify candidate genes associated with the desired phenotype.
Though high-throughput screening is inherently complex, it follows a well-defined workflow—from meticulous library design and amplification, to precise viral packaging and cell transduction, and finally to phenotypic evaluation and sequencing-based data analysis. Depending on the research objective, screening strategies can be flexibly adapted to include:
These technical modalities enable high-specificity investigations into both gain- and loss-of-function gene phenotypes. In more advanced applications, we also support integration with specialized screening methods, including:
To facilitate the implementation of these workflows, Creative Biogene provides detailed experimental protocols and design support.
Creative Biogene is committed to delivering top-tier sgRNA library services globally. With expert teams and robust infrastructure, we meet diverse research and clinical needs. Partner with Creative Biogene to advance CRISPR-based innovations in life sciences and health.
Q1: How do you ensure sgRNA specificity and minimize off-target effects?
A1: We employ multiple strategies: (1) Bioinformatics prediction and evaluation; (2) GC content and sequence length control; (3) Multiple sgRNAs per gene; (4) Whole-genome alignment to identify potential off-targets.
Q2: What vector systems are available?
A2: We primarily offer lentiviral vectors (e.g., LentiCRISPRv2) known for high transduction efficiency and broad applicability. Custom vectors are also available.
Q3: How is library quality ensured?
A3: We conduct comprehensive QC: (1) Sanger validation; (2) NGS-based coverage and uniformity assessment; (3) Strict transformation and amplification controls; (4) sgRNA representation analysis; (5) Detailed reports.
Q4: Are off-the-shelf libraries available?
A4: Yes. We offer genome-wide knockout libraries for human and mouse models, as well as specialized libraries targeting specific pathways and diseases.
Q5: What should be considered for library screening experiments?
A5: Key factors include: (1) Sufficient MOI for single-copy integration; (2) Maintaining library complexity; (3) Proper controls; (4) Optimized screening pressure and timepoints; (5) Replicate experiments.
Q6: What does post-screening data analysis include?
A6: Analysis includes: (1) Sequencing QC; (2) sgRNA abundance profiling; (3) Statistical significance testing; (4) Functional enrichment and pathway analysis; (5) Network construction and visualization. We provide full bioinformatics support.
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