Reporter genes are important for knock-in strategies. For the introduction of foreign DNA, both the selected cDNA and the reporter gene are needed to be brought into the genome, where the reporter gene acts as a selection marker for positive clones. Another interesting application of reporter gene knocked-in technology is not only to produce a null mouse but also to help track the targeted gene's expression pattern through the mutated gene. Creative Biolabs has successfully applied the reporter/tag gene knock-in technologies in the generation of models of different applications. We offer customized model creation services for our clients to meet their exact research needs.

Introduction of Reporter/Tag Gene

Genes that are chosen as reporters usually carry visually identifiable characteristics. A type of reporter genes can encode fluorescent and luminescent proteins such as green fluorescent protein (GFP) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range, and the enzyme luciferase, which catalyzes a reaction with luciferin to produce light. Besides, the E. coli lacZ gene is another commonly used reporter, which encodes the protein beta-galactosidase thus causing bacteria expressing the gene to appear blue when grown on a medium that contains substrate analog X-gal. In addition, marker genes, which are capable of conferring resistance to certain antibiotics, can be knocked-in for indication of successful introduction of foreign genetic material.

Applications of Reporter/Tag Mouse Models

  • For studying the transcriptional activity of gene promoter - the targeted gene coding sequence is replaced with a reporter gene coding sequence (e.g., EGFP), where the endogenous gene is knocked out as the reporter gene is simultaneously knocked-in via homologous recombination. Upon recombination, the promoter of the targeted gene will drive expression of the inserted reporter gene.
  • For monitoring the expression, localization, and transportation of proteins in vivo - the visualization of a gene's expression can be realized by adding a tag sequence into the end (or the beginning) of the coding sequence of the targeted gene which will result in a fusion protein for visualizing gene expression.
  • For target identification and preclinical evaluation of drug candidates

Features of Reporter/Tag Gene Knock-in Models

Reporter gene mouse models have proven to be powerful tools for studying the transcriptional activity of a specific gene promoter, monitoring the protein expression and protein trafficking, and for the screening of drug candidates. Besides, reporter mouse models provided by Creative Biolabs are highly sensitive and allows one to follow the response of the same animals over time, providing a kinetic readout in vivo. However, the targeted gene can be knocked-out or disrupted with the reporter gene knocked-in. To prevent this from happening, an internal ribosome entry site (IRES) can be used to co-express both the target gene and the reporter gene (see Figure 1).

How to Generate a Reporter/Tag Mouse Model?

Knock-in models can be produced with either ESC/HR-Based Gene Targeting Technology or CRISPR/Cas9-Based Targeting Strategy. For the former, knock-in mutant is realized by homologous recombination in embryonic stem cells and subsequent injection of modified embryonic stem cells in preimplantation stage mouse embryos. For the latter, potential knock-in mouse models can be obtained with the injection of CRISPR/Cas9 components (sgRNA, Cas9mRNA with or without a repair DNA) into the mouse embryo.

In the meanwhile, Creative Biolabs offers other types of knock-in models that you may be interested in:

At Creative Biolabs, our technical team is expert in the assessment of project feasibility and is willing to assist you in the right selection of the targeting strategy that would best meet your project goals. Contact or inquire us to discuss the ideal custom mouse model to boost the impact of your next project and beyond.

For Research Use Only.



Online Inquiry
Name:
*Phone:
*E-mail Address:
*Service & Products Interested:
Project Description:
Contact Us USA

Tel:
Fax:
Email:
UK

Tel:
Email:

Germany

Tel:
Email:

Follow us on:
Copyright © 2024 Creative Biolabs.