Gene Knock-in Services

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Gene knock-in services offer a powerful and precise approach for genetic modification, enabling the insertion of specific DNA sequences into the genome. This technique is invaluable for studying gene function, creating disease models, and developing innovative therapies. By introducing new genetic material into a targeted location within the genome, researchers can gain insights into gene expression, protein function, and regulatory mechanisms. Gene knock-in is a cornerstone of modern genetic research and therapeutic development.

Generation of pRSAD2 gene knock-in pig via CRISPR/Cas9 technology (Z Xie, et al.,2020)

Highlight:

Precision and Accuracy: Utilizing advanced CRISPR/Cas9 and other gene-editing technologies to ensure precise and accurate insertion of genes at target locations within the genome.

High Efficiency: Achieving high knock-in efficiency with optimized protocols, reducing the time and resources required for successful gene editing.

Quality Assurance: Implementing rigorous quality control measures throughout the process, including sequence verification and functional assays, to ensure the integrity and functionality of the inserted genes.

Scalable Services: Capable of handling projects of various scales, from small pilot studies to large-scale production, catering to both academic and commercial research needs.

Overview Service Process Examples and Solutions Applications Frequently Asked Questions

Overview

Gene knock-in involves the precise insertion of a gene or other DNA sequence into a specific locus within the genome. This is typically achieved using advanced gene-editing technologies such as CRISPR/Cas9, TALENs (Transcription Activator-Like Effector Nucleases), or zinc finger nucleases. These tools create targeted double-strand breaks at the desired genomic location. A donor template carrying the desired sequence, flanked by homologous regions to the target site, is then introduced. The cell's natural homology-directed repair (HDR) mechanism integrates the new sequence into the genome, enabling the study of its function and effects.

Service Process

The process of gene knock-in involves several critical and interrelated steps:

Target Identification: Selecting the specific genomic locus for the knock-in based on the research objective or therapeutic goal. Accurate target identification is crucial for the success of the knock-in process.
Donor Template Design: Designing a donor template that carries the desired DNA sequence flanked by regions homologous to the target site. This template guides the integration of the new sequence during the repair process.
Guide RNA Design: For CRISPR/Cas9, designing guide RNAs that direct the nuclease to the precise genomic location. This step ensures specificity and accuracy in creating the double-strand break.
Delivery of Editing Components: Introducing the gene-editing tool, guide RNA, and donor template into the target cells using methods such as electroporation, viral vectors, or lipid nanoparticles. Efficient delivery is vital for successful HDR.
Selection of Knock-in Cells: Isolating cells that have successfully incorporated the new sequence. This selection process may involve the use of selectable markers or advanced sorting techniques to enrich the population of knock-in cells.
Validation: Confirming the successful integration of the new sequence through sequencing and functional assays. This step ensures that the knock-in is precise and that the cells exhibit the expected phenotypic changes.

For more information about our Gene Knock-in Services or to discuss your specific needs, please contact us. Our team of experts is available to provide guidance and support for your research projects, ensuring you achieve your scientific and therapeutic goals.

Examples and Solutions

The following table provides an overview of various case studies in gene knock-in and the solutions we offer to support your research and therapeutic endeavors:

Case Study	Description	Solutions We Offer
Gene Therapy for Hemophilia	Inserting functional F8 gene into liver cells to correct clotting deficiency.	CRISPR/Cas9 HDR, donor template design, cell therapy preparation, and clinical trial support.
Oncology Research	Introducing oncogenes or tumor suppressor genes into cell lines to study cancer.	Gene knock-in using CRISPR/Cas9, validation assays, and cancer modeling.
Neurodegenerative Disease Models	Knock-in of mutant genes associated with Alzheimer's into neural stem cells.	CRISPR/Cas9 HDR, donor template design, neural differentiation, and functional assays.
Cardiovascular Disease Studies	Inserting genes involved in heart disease into cardiac progenitor cells.	TALENs/CRISPR/Cas9 HDR, donor template design, cardiac cell differentiation, and phenotype analysis.
Immunology Research	Introducing cytokine genes into immune cells to enhance immune response.	Gene knock-in tools, immune cell assays, and functional testing.
Metabolic Disorder Research	Inserting genes involved in metabolic pathways into liver cells.	CRISPR/Cas9 HDR, donor template design, metabolic profiling, and functional analysis.

Applications

The applications of gene knock-in are extensive and transformative, including:

Functional Genomics: Studying the roles of specific genes and regulatory elements by inserting reporter genes, tags, or regulatory sequences.
Disease Modeling: Creating accurate cellular and animal models of human diseases by introducing disease-associated mutations or genes.
Gene Therapy Development: Inserting therapeutic genes into specific loci to correct genetic disorders or enhance cell function.
Protein Function Studies: Investigating protein function by inserting epitope tags or fluorescent markers for visualization and tracking.
Drug Development: Testing the effects of new genes or regulatory elements on cellular pathways to identify potential therapeutic targets.

Frequently Asked Questions

Q: What is gene knock-in?

A: Gene knock-in involves the targeted insertion of a gene or other DNA sequence into a specific locus within the genome to study gene function, create disease models, and develop therapeutic strategies.

Q: How is gene knock-in performed?

A: Gene knock-in is performed using advanced gene-editing technologies such as CRISPR/Cas9, TALENs, or zinc finger nucleases. These tools create double-strand breaks at the desired genomic location, and a donor template with the desired sequence is integrated into the genome via homology-directed repair.

Q: What are the applications of gene knock-in?

A: Applications include functional genomics, disease modeling, gene therapy development, protein function studies, and drug development. Each application leverages the precise insertion of DNA sequences to study and manipulate gene function and expression.

Q: What are the key steps in the gene knock-in process?

A: Key steps include target identification, donor template design, guide RNA design (for CRISPR/Cas9), delivery of editing components into cells, selection of successfully knocked-in cells, and validation of genetic modifications through sequencing and functional assays.

Q: Why is gene knock-in important?

A: Gene knock-in is important for understanding gene function, studying disease mechanisms, developing gene therapies, investigating protein functions, and advancing drug discovery. It provides precise and reliable tools for genetic research and therapeutic development.

Please note that all services are for research use only. Not intended for any clinical use.

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