Gene Editing Technology

What types of gene editing technologies are there? What are the advantages of the CRISPR/Cas9 gene editing technology? What is embryonic stem cell targeting (ESC targeting)?

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To provide our customers with the ideal animal model that is in line with their requirements from basic research to drug development, Shanghai Model Organisms Center (SMOC) has always focused on gene editing and model development of model organisms. We have a professional project design team, an AI-driven automated analytical system known as SmartEddi, as well as an experienced and highly-trained research and technical team. Since 2000, we have established a reliable and efficient service platform for genetically engineered mice and we are committed to providing the best-in-class solutions for genetically engineered animal models.


The following three types of genetic engineering technologies are used to generate genetically engineered mouse(GEM) models:



Comparison

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Embryonic Stem Cell Targeting


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In murine embryonic stem cells, the exchange of an endogenous allele of a target gene for a mutated copy via homologous recombination allows the generation of  targeted ES cells with defined mutations in their genome  that was previously designed by the researcher.Genetically engineered ES cells remain pluripotent and are able to develop into germ cells in the chimeric animals, thereby ensuring the germline transmission which results in a genetically engineered mouse model.As of today, this remains the most classic and reliable genetic engineering or gene editing technology for mouse models.


Currently, SMOC provides mouse ES cells with three types of genetic background: C57BL/6, 129/S6, B6;129.

These mouse ES cells can be used to generate the following types of mouse models:

  • Gene knockout

  • Conditional gene knockout

  • KO first

  • Gene knock-in

  • Point mutation

  • Conditional point mutation

  • Targeted gene overexpression

  • Humanization


SMOC will analyze and evaluate each project, combining time and risk factors to select the most appropriate technology (ESC targeting or CRISPR gene editing).

Contact us to discuss how to use ES cell targeting technology to obtain your animal model with our technical consultants of SMOC.

General procedure for obtaining mouse models using ESC targeting technology

  • Design and generate homologous recombination vectors

  • Transfer the homologous recombination vectors into murine ES cells of mouse

  • Screen and validate the positive targeted ESC clones

  • Microinject positive ESC clones into mouse blastocysts

  • Transplant the injected mouse blastocysts into the uterus of pseudopregnant female mice

  • Generate and screen positive chimeric mice F0

  • Obtain F1 heterozygous mice by mating positive F0 with wild-type mice or FLP mice


CRISPR Gene Editing Technology


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Advantages of CRISPR gene editing technology

  • The R&D cycle is greatly shortened compared with the traditional gene targeting method.

  • Overcomes the limitations of mouse genetic strains and achieves genetic editing on different genetic backgrounds or based on existing genetically engineered mouse models.

  • Reduces the cost of customization for knockout and knock-in mouse models.


SMOC uses the latest CRISPR/Cas9 gene editing technology to generate your custom genetically engineered mouse model.

SMOC will analyze and evaluate each project, combining time and risk factors to select the most appropriate technology (CRISPR gene editing or ES cell targeting).

Contact us to discuss how to use CRISPR technology to obtain your animal model with the technical consultant of SMOC.

General procedure for obtaining mouse models using CRISPR gene editing technology

  • Select target site and design sgRNA, design homologous recombination vector (optional)

  • Prepare sgRNA and Cas9 mRNA, generate homologous recombination vector (optional)

  • Microinject sgRNA and Cas9 mRNA (and homologous recombination vector (optional)) into mouse fertilized eggs

  • Transplant fertilized eggs after microinjection into the oviducts of pseudopregnant female mice

  • Generate and screen positive chimeric mice F0

  • Obtain F1 heterozygous mice by mating positive F0 with wild-type mice


Random Transgenes

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A pre-Designed gene (or genes) are randomly integrated into the mouse genome through pronuclear microinjection to generate transgenic mice. Transgenic mouse models can also be obtained more efficiently using the piggyBac transposon system.


General procedure for obtaining random transgene mouse models

  • Design and generate transgenic plasmid

  • Inject linearized transgenic plasmid fragments into mouse fertilized eggs

  • Transplant fertilized eggs after injection into the oviducts of pseudopregnant female mice

  • Generate and screen positive chimeric mice F0

General procedure for obtaining transgenic mouse models using the piggyBac transposon system

  • Design and generate PiggyBAC transposon plasmid

  • Inject transposon plasmid and PiggyBAC transposase into mouse fertilized eggs

  • Transplant fertilized eggs after injection into the oviducts of pseudopregnant female mice

  • Generate and screen positive chimeric mice F0


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