Overview

Genes play a key role in the development of obesity. A growing number of studies have revealed genetic abnormalities associated with human obesity. In-depth understanding of the effects of these genetic variants on physiological processes such as energy metabolism, fat storage, and appetite regulation, and evaluating whether they are used as potential therapeutic targets, is crucial in obesity research.

Targeted Gene Knockin: Unlocking the Genetic Basis of Obesity

Protheragen provides a high-quality, customized obesity gene knock-in service to help researchers achieve breakthroughs in obesity mechanism research, drug target validation, and efficacy evaluation. Knock-in models integrate exogenous gene fragments into specific sites in the genome to better analyze their functions and study related physiological changes. Our obesity knock-in program covers all aspects of the process and offers a wide range of options to best meet our clients' needs.

Project Consultation and Design

We gain a deep understanding of your research needs and gene editing goals, and evaluate the entire project based on relevant information systems to develop optimal knock-in strategies, sgRNAs, and donor templates. When designing sgRNAs, we utilize online tools to predict targeting efficiency and off-target effects, and select sgRNA sequences with the lowest off-target risk and highest targeting efficiency.

Knock-in and Cell Processing

Transfection of editing tools into cells (fertilized eggs and other specific cell lines) is performed by a variety of methods (electroporation, transfection, viral transduction, etc.). After screening and single-cell cloning, we obtain knock-in stable cell lines. Knock-in animal models also involve embryo transfer, priming, line expansion, and other manipulations.

Customizable Knock-in Types:

• Site-specific mutation knock-in
• Reporter gene/tag protein knock-in
• Humanized knock-in
• Conditional knock-in
• Large DNA fragment knock-in

QC Validation

We validate knock-in and analyze off-target effects by PCR, Sanger sequencing, etc., and detect relevant protein expression by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blot, fluorescence-activated cell sorting (FACS), etc. We also analyze knock-in and off-target effects by PCR and Sanger sequencing. In addition to this, analyses such as sterility testing and chromosome analysis are also selected.

Downstream Functional Validation

We carry out many types of phenotyping and functional validation, including but not limited to cytotoxicity, cell viability, cell proliferation, and animal phenotyping (glucose tolerance, insulin sensitivity, inflammatory factor assays, etc.). Various assays are flexibly selected.

Workflow

Applications

• Knocking in specific genes and analyzing the phenotypic changes in animals provides valid information for screening potential anti-obesity therapeutic targets.
• Specific knock-in models are used to evaluate the efficacy and potential toxicity of drugs targeting the gene or related pathways.
• Accurately modeling genetic changes by knock-in to study their functional roles at the cellular, animal levels.

Advantages

• Our team has many years of experience in constructing gene editing models, especially in the field of obesity, and can flexibly choose the optimal solution according to the project requirements.
• We significantly improve the success rate of gene knock-in and ensure the genetic stability of the model through fine design and strict process control.
• We provide end-to-end services from pre-consultation, protocol design, project execution, to post-project reporting and technical support.

One-stop Anti-obesity Research Support

Publication Data

Frequently Asked Questions

1. What information do I need to provide to start the project?

   You need to provide the name of the gene you are interested in, the species, the region of the gene targeted, the specific type of gene editing you wish to achieve (e.g., point mutation, inserted sequences, reporter genes, etc.), and relevant research background information.

2. How do you ensure the accuracy of the gene knock-in?

   QC validation is a key component in the construction of knock-in models. We use multiple validation methods, including but not limited to genomic PCR, Sanger sequencing, off-target effect assay, chromosome analysis, protein expression analysis, etc. In addition, we also perform detailed tests on cell activity and animal phenotype.

Protheragen focuses on obesity research needs, masters cutting-edge gene editing technologies, and provides professional technical support and project services. Welcome to contact us to communicate any project and help you quickly obtain key research tools to accelerate the research process.

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.

• Candidate Obesity Gene Screening Service
• Obesity Gene Knockout Service
• Obesity Gene Overexpression Service
• Obesity Gene Silencing Service
• Obesity Multigene Gene Editing Service
• Obesity Single Gene Editing Service