Target Genome Editing Service

Definition of Target Genome Editing

Gene Knockout Platforms Used for Gene Knockout Process of CRISPER Gene Knockout

Gene Overexpression Technologies Used for Gene Overexpression Applications for Gene Overexpression

Relevant Services FAQ Resources

With decades of experience, Creative Biolabs has established itself as a leader in the field of target genome editing, a transformative approach in genetic research that allows precise modifications to DNA sequences. Scientists in Creative Biolabs utilize the advanced CRISPR-based system as a genome-editing tool to knock out or silence specific genes, modulate gene expression, or upregulate genes to enhance protein production. This cutting-edge system contributes to the targeted modification of gene expression, facilitating the screening of potential therapeutic targets, and enabling the study of how genomic alterations may impact organisms. The versatility of CRISPR-based system makes it ideal for various applications, from basic research to drug discovery and therapeutic development.

What is Target Genome Editing?

Target genome editing refers to the process of precisely altering specific sequences in an organism's DNA to understand gene function, model diseases, or develop new therapeutic strategies. Researchers can induce targeted mutations or edits in specific genes by employing a range of engineered nucleases such as the CRISPR-based system and transcription activator-like effector nuclease.

Gene Knockout

Gene knockout involves deleting or inactivating a specific gene to assess its function. This is typically achieved by inducing double-strand breaks (DSBs) in the DNA at the target site, followed by repair via non-homologous end joining (NHEJ), which often introduces insertions or deletions (indels) that disrupt the gene's coding sequence. This results in a complete loss of function in the targeted gene.

At Creative Biolabs, CRISPR technology has been fully integrated into our gene editing platform, offering a powerful tool for gene knockout studies. The CRISPR-based system is engineered to disrupt genes efficiently, facilitating the creation of isogenic cell lines and animal models. Our optimized protocols minimize off-target effects while maximizing editing efficiency.

Platforms Used for Gene Knockout

CRISPR-based System: The most widely used system for gene knockout, CRISPR-based system, utilizes a single-guide RNA (sgRNA) to direct the Cas9 enzyme to the specific DNA sequence. Once bound, Cas9 introduces DSBs at the target site, leading to gene disruption through NHEJ.
Transcription Activator-Like Effector Nucleases: Transcription activator-like effector nuclease employs customizable DNA-binding proteins fused with a nuclease (FokI) to introduce breaks at target sequences. Although highly specific, they are less user-friendly compared to CRISPR-based system.

The types of our genetic manipulation techniques including but not limited to:

Platforms	Advantages	Conditions
CRISPR-basedSystem	Small guide RNA serves as the recruiter Highly versatile and easy to design High-efficiency andcost-effective High Throughput Screening Potentially adaptable for almost any gene disruption	Best suited for rapid, large-scale knockouts in various organisms and cell lines. Suitable where off-target effects can be managed, especially in simpler genomic environments.
Transcription Activator-like Effector Nuclease	Highly specific and low off-target effects Programmable with modular arrays Each module recognizes a single base pair Effective for targeted modifications with greater accuracy than CRISPR-based system	Ideal for projects requiring high precision in complex genomes or gene-rich regions. Suitable where off-target effects must be minimized, especially in delicate or medically significant genomic areas.

CRISPR Gene Knockout Workflow

Our CRISPR-based gene knockout process follows a streamlined workflow, ensuring high precision and reproducibility in genomic alterations:

Design and Synthesis of gRNA

Our experts design the optimal guide RNA (gRNA) that targets the specific gene sequence. This is followed by gRNA synthesis.

CRISPR-based System Delivery

The Cas9 nuclease and gRNA are introduced into the cells via electroporation, viral transduction, or chemical transfection, depending on the cell type.

Targeted Gene Editing

Once inside the cells, Cas9 introduces a double-strand break at the target site, leading to gene knockout via NHEJ.

Screening and Validation

After editing, the cells are screened to identify those that carry the desired gene disruption. This is confirmed by sequencing, Western blotting, or PCR.

Functional Analysis

The knockout cells are subjected to functional assays to study the effects of the disrupted gene on cellular processes.

Gene Overexpression

Gene overexpression aims to increase the expression of a specific gene to study its effects on cellular processes. This approach is crucial for understanding the impact of gene dosage on phenotypes and can be used in disease models or biotechnological applications such as enhanced protein production.

Technologies Used for Gene Overexpression

CRISPRa (CRISPR Activation): A modified version of the CRISPR system, CRISPRa uses a catalytically dead Cas9 (dCas9) fused with transcriptional activators to upregulate gene expression. This tool is highly versatile, as it can be targeted to any gene promoter.
Viral Vectors: Lentiviral or adeno-associated viral (AAV) vectors are often employed to introduce an exogenous gene or activate an endogenous gene. They integrate into the genome or exist as episomes in the host cell, leading to stable or transient gene overexpression.
Transcription Activator-Like Effectors (TALEs): Similar to transcription activator-like effector nuclease, TALEs can be modified to serve as transcriptional activators that enhance the expression of a target gene without introducing DSBs.

Technology	Advantages	Conditions
CRISPRa (CRISPR Activation)	- Highly flexible, reversible control over gene expression.	- Suitable for precise, reversible gene activation without altering the DNA sequence, ideal for studying gene function in model organisms.
Viral Vectors	- Efficient and stable long-term overexpression.	- Ideal for stable overexpression in vivo and in vitro where prolonged gene expression is needed, especially for therapeutic studies.
TALE Transcription Activators	- Highly specific with minimal off-target effects.	- Ideal for precise overexpression in complex genomes with overlapping regulatory elements, particularly useful in tightly regulated gene systems.

Applications for Gene Overexpression

Creative Biolabs offers gene overexpression services to support a wide range of scientific research, including the following applications:

Heterologous Expression: Overexpressing genes in different organisms allows for functional studies in alternative model systems, expanding the scope of biological research.
Drug Target Identification: Overexpressing genes help uncover target protein functions in drug interactions.
Pathway Analysis: Wild-type protein overexpression in mutants reveals drug-related pathways and compensatory mechanisms.
Epistasis Testing: Overexpression aids in epistasis tests, revealing genetic interactions and pathway hierarchies.
Mutation Reversion: Mutant protein overexpression restores functionality in deficient cells for functional studies.
Loss-of-Function Substitute: Overexpression generates mutations when loss-of-function studies are impractical.
Interaction Mapping: Overexpression outputs assist in mapping protein-protein interaction networks.
Modifier Screens: Overexpression identifies genetic modifiers that alter specific phenotypes.
Heterologous Expression: Overexpression in different organisms enables functional studies across species.

As a leader in target genome editing, Creative Biolabs continues to push the boundaries of gene manipulation for research and therapeutic purposes. From gene knockout to overexpression, our platform leverages the latest technologies, including CRISPR-based system, to offer precision and reliability. Whether in disease modeling, drug discovery, or agricultural applications, Creative Biolabs ensures that every project benefits from our expertise and state-of-the-art methodologies . Please do not hesitate to contact us for more details.

Relevant Services

Gene Conditional Overexpression Gene Conditional Overexpression is a precise method that allows the controlled activation or overexpression of a specific gene under predefined conditions, typically in response to an external stimulus such as a chemical inducer, temperature shift, or light. This system provides researchers with the ability to control the timing, location, and intensity of gene expression, making it a highly valuable tool in various scientific and biomedical applications. Creative Biolabs utilizes this technique to enable regulated overexpression of genes in diploid organisms. Combine conditional gene expression with tailored regulatory systems to provide a reproducible and reliable approach to studying gene essentiality and function. It is especially valuable for large-scale gene identification and validation in pathogenic microorganisms, allowing for dynamic control over gene activity in various biological systems.

Gene Silencing Creative Biolabs offers a comprehensive RNA interference (RNAi) service platform, enabling effective gene silencing for various research applications. Our services include:

Custom design and synthesis of siRNA or shRNA.
Construction of siRNA or shRNA expression vectors.
siRNA or shRNA transfection services.
Validation of gene silencing through mRNA quantification via RT-PCR and protein expression analysis via Western Blot.

FAQs

What is the difference between gene knockout and gene silencing?

Gene knockout completely disrupts a gene's function by introducing a mutation or deletion, while gene silencing reduces or inhibits gene expression without altering the underlying DNA sequence. Both methods are used to study gene function but operate through different mechanisms.

How does Creative Biolabs ensure high specificity in CRISPR-based system genome editing?

Creative Biolabs employs advanced guide RNA design tools and optimized protocols to target specific DNA sequences accurately, minimizing off-target effects. Our rigorous validation steps, including sequencing and protein assays, further ensure the precision of each genome editing project.

What are the advantages of CRISPR-based system over transcription activator-like effector nuclease for gene knockout?

CRISPR-based system is more user-friendly, allowing rapid and flexible design for gene editing with higher efficiency and lower cost compared to transcription activator-like effector nuclease. It also enables simultaneous editing of multiple genes, which is not easily achievable with other technologies.

What applications can genome editing be used for in drug discovery?

Genome editing is widely used in drug discovery to identify and validate drug targets, study disease mechanisms, and model genetic mutations in cellular or animal systems. This approach helps accelerate the development of novel therapeutic interventions.

How does Creative Biolabs validate successful gene knockouts or overexpression experiments?

Creative Biolabs validates successful gene knockouts and overexpression using a combination of DNA sequencing, qPCR, and protein expression analysis. These methods ensure that the intended genetic modifications have been achieved and are functioning as expected.

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