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Tissue-specific Promoter Driven SSR System
Site-specific recombinases (SSRs) are DNA modifying enzymes that recognizing specific target sites. Generally, the SSRs catalyze recombination reactions of genomic DNA according to the location and orientation of target sites, including delete, integrate, invert and translocate.
What's Tissue-specific Promoter Driven SSR System?
The tissue-specific SSR is created by fusing the recombinase coding sequence to a promoter that drives gene expression specifically in the desired tissue or cell type. When the system is expressed in cells of the target tissue, it causes a specific and efficient recombination of the specific sites and subsequent modification of the target gene. Tissue-specific promotor driven SSR systems are very powerful tools for figuring out the gene function and the mechanism of its regulation. By utilizing cell/tissue-specific regulatory elements (promotors and enhancers), exogenous inducers (tamoxifen and tetracycline), or other delivery methods (such as viral vectors, extracellular vesicles, et al.), the systems are able to achieve conditional KO in tissues or organs of interest.
Principle of Tissue-Specific KO via SSR System
Generally, the strategy of generating tissue-specific KO mouse via SSR systems includes several steps. Here we take Cre/loxP system as an example:
- Generating mice that are heterozygous for a loxP-flanked allele (floxed mouse) and heterozygous for the tissue-specific promoter driven Cre transgene.
- Crossbreeding the floxed mouse with the Cre mouse, and 50% of the offspring will be heterozygous for both loxP and Cre.
- Backcrossing the offspring to the floxed parent mouse, 25% of the offspring will be the tissue-specific OK mouse.
What Can Tissue-specific Promoter Driven SSR System Do?
- Study gene function: By creating tissue-specific knockouts or knock-ins of specific genes, researchers can study the function of those genes in a particular tissue and how their loss or alteration affects the tissue's development, function, and disease susceptibility.
- Create animal models of human diseases: By using tissue-specific promoter-driven SSR, researchers can create animal models of human diseases, allowing them to study the underlying mechanisms of the diseases and test potential therapies.
- Study cell differentiation: By targeting SSR to different cell types during development, researchers can study the differentiation and maturation of those cells and the role of specific genes in those processes.
- Improve gene therapy: By using SSR to precisely and specifically target and modify therapeutic genes, researchers can improve the efficiency and specificity of gene therapy for various diseases.
Types of Tissue-specific Promoters and Their Applications
Due to the great value of SSR systems as genetic engineering tools, many tissue/organ-specific promoter/enhancer of SSR driver have been developed. The various tissues are mainly divided into 6 physiological systems: including nervous, integumentary, immune, musculoskeletal, digestive and urogenital system.
Nervous System
Nervous system mainly consists of two parts: the central nervous system (CNS) and the peripheral nervous system (PNS). The Cre-loxP system is widely used for neurological research, including generating conditional KO models, brainbow and many others.
Integumentary System
The integumentary system consists of skin and its appendages such as hair, nails, sebaceous glands and sweat glands. Cre-loxP system are usually used for gene functional study.
Immune System
Tissue-specific SSR system for immune system is widely used for gene functional study of immune system, as well as the lineage tracing of immune cells, such as macrophage, dendritic cells, T and B cells, haematopoietic cells, et al.
Musculoskeletal System
Tissue-specific SSR systems are usually employed as tools for gene functional studies, as well as musculoskeletal disease, such as vascular diseases, cartilage-related diseases, and others.
Digestive System
Digestive system includes digestive tract and various auxiliary organs. Usually, the tissue-specific promoter driven Cre-loxP system are used for studying glandular development and related diseases.
Urogenital System
This system consists of the urinary system, such as the bladder and kidneys, and the reproductive organs, such as the ovaries and testes. The Cre-loxP system serves as a tool for gene functional study, embryo differentiation and spermatogenesis.
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