Application of reagent for detecting or inhibiting circ7598
A technology of reagents and preparations, which is applied in the application field of detection or inhibition of circ_7598 reagents, can solve problems such as inability to achieve long-term remission or complete cure, affect patients' physical and mental health and quality of life, and high recurrence rate, so as to slow down cell proliferation and cell proliferation. The effect of the cycle process
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Embodiment 1
[0042] Example 1: The real existence verification of circ_7598
[0043] 1. Circ_7598 circularization site sequence verification
[0044] Divergent primer and convergent primer were designed by Oligo 7 software to amplify the circularization site of circ_7598 and the parental gene sequence, respectively. Take the cDNA and gDNA of skin tissue or HaCaT cells (human immortalized keratinocytes, purchased from the National Biomedical Experimental Cell Resource Bank) as templates, and use the above primers to carry out RT-PCR reaction and PCR product agarose electrophoresis, such as figure 1 The electrophoresis results of B show that: using cDNA or gDNA as the template, the convergent primer amplifies the target fragment with the expected size, while the divergent primer only uses cDNA as the template to amplify the target fragment with the expected size. Further use Sanger The target fragment was sequenced by the next generation method, and the sequencing result was consistent with...
Embodiment 2
[0047] Example 2: Cell localization study of circ_7598
[0048] In order to clarify the localization of circ_7598 in keratinocytes, the cytoplasmic and nuclear components of HaCaT cells were separated, RNA was extracted, GAPDH was used as the cytoplasmic reference, and U6 was used as the nuclear reference, and the expression of circ_7598 was detected by real-time fluorescence quantitative PCR reaction. The detection results showed that circ_7598 was mainly located in the cytoplasm ( image 3 ). In addition, the RNA in situ hybridization kit (F16501 / 50) of Shanghai Gema Gene Co., Ltd. was also used, and experiments were carried out according to the instructions to visualize the localization of circ_7598, such as Figure 4 Shown: The red fluorescence of labeled circ_7598 is wrapped around the nuclear-stained blue fluorescence.
Embodiment 3
[0049] Example 3: Expression of circ_7598 in psoriasis tissues and cells
[0050] 1. 40 cases of psoriasis skin tissue and 40 cases of normal skin tissue were collected from the Department of Dermatology of the Second Xiangya Hospital of Central South University, of which 20 cases of psoriasis skin tissue and 20 cases of normal skin tissue were stored in liquid nitrogen for preservation. To explore the expression differences of circ_7598 in the full-thickness skin of the two groups of samples; after the remaining 20 samples in each group were separated from the dermis, the epidermal tissues were collected to explore the expression differences of circ_7598 in the epidermal tissues of the two groups of samples. All patients were diagnosed with psoriasis vulgaris by skin histopathological examination in our hospital, and other skin diseases such as systemic lupus erythematosus, atopic dermatitis, eczema, neurodermatitis were excluded. Topical glucocorticoids, retinoids, vitamin D...
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