With the deepening of neuroscience research, scientists need to pay attention to many different types of neurons or glial cells, such as a variety of GABAergic interneurons and microglia. Unfortunately, these cells often do not have mature, non-leaking promoters to choose from. Therefore, it is very important to achieve cell type-specific labeling or manipulation. Recombinase systems such as Cre-LoxP or Flp-FRT are ideal tools for labeling specific cell types.
To put it simply, this type of recombinase system uses site-specific recombinases (SSRs) to mediate recombination between Recombination tar-get sites (RTs) to achieve specific sites manipulation, such as gene knockout, gene insertion, gene flip, and gene translocation. Since this technology can effectively overcome the shortcomings of non-specificity or low recombination efficiency, it has gradually occupied a dominant position in the field of functional gene research in recent years.
Cre recombinase not only has catalytic activity but is similar to restriction enzymes. It can recognize specific DNA sequences, namely loxP sites so that gene recombination occurs between two loxP sites. Cre recombinase does not require any cofactors, and can act on DNA substrates of various structures, such as linear, circular, and even supercoiled DNA. In addition, the activity of Cre recombinase is controllable, so that it can be expressed only in a certain type of cell type, or it can be triggered by specific external stimuli (such as chemical signals, thermal stimuli). In the field of neuroscience, this technology can be used for specific labeling of neural circuits, functional research of endogenous genes in specific cell types, and construction of mouse models.
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