Mouse Shank 3 gene cRNA probe and in-situ hybridization color development method

Abstract

The invention provides a mouse Shank 3 gene cRNA probe and an in-situ hybridization color development method. The cRNA probe comprises a nucleotide sequence with complementary sites 3465-4108 of a reference sequence NM_021423.3 of mouse Shank 3 gene mRNA. The in-situ hybridization color development method comprises the following steps: putting a mouse brain tissue sample in a hybridization solution; after hybridization, rinsing, closing and combining with an antibody; amplifying with TSA-biotin and performing fluorescent color development. The cRNA probe provided by the invention has high specificity on the Shank 3 gene, can efficiently finish recognition of the Shank 3 gene in the brain tissues of different individuals, can ensure the specificity and high efficiency of in-situ hybridization color development of the mouse Shank 3 gene, realizes a better color development effect and can be combined with other hybridization or immunofluorescent staining.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a Shank 3 gene cRNA probe and a fluorescence in situ hybridization color development method thereof. Background technique [0002] The Shank family is a newly discovered scaffold protein, which can interact with various proteins in cells through different structural domains and play multiple roles. Studies have shown that the Shank3 gene is closely related to the onset of autism, and mutations in the Shank3 gene can lead to autism-like behaviors. Studies such as protein interaction have shown that Shank3 protein may interact with various intracellular proteins through different structural domains, and ultimately affect the structure and function of the cytoskeleton and synapses. However, the morphological evidence in this regard is obviously insufficient, mainly due to the fact that none of the commercially available antibodies can clearly show the intracellular localizati...

AI Technical Summary

Problems solved by technology

However, screening cRNA probes with both hybridization specificity and chromogenic sensitivity for the Shank family, especially the Shank3 gene, is still a difficult problem in the field

Although the use of reagents with the function of amplifying the color development effect is more conducive to color development, there is no amplification system for cRNA probe in situ hybridization color development suitable for the Shank family, especially Shank3, which also leads to the corresponding An important reason why cRNA probes have not been obtained

In addition, the existing cRNA probe-based chromogenic methods have disadvantages such as complex composition of chromogenic reagents (such as amplification reagents, hybridization solutions), and at the same time, the chromogenic effect still needs to be improved

[0004] So far, there are no reports on cRNA probes and fluorescent in situ hybridization methods for Shank3 gene

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