兔VX2細胞的核酸適配體篩選和結構優化
首發時間:2022-05-07
摘要:目的:篩選兔VX2細胞的核酸適配體,并對獲得的核酸適配體進行結構優化和性能考察。方法:利用Cell-SELEX技術,以兔VX2細胞為靶細胞,兔腎RK13細胞為對照細胞進行核酸適配體的篩選;對獲得的核酸適配體進行序列的裁剪、替換等處理后對其進行性能考察。結果:經過12輪的篩選獲得了5條兔VX2細胞的核酸適配體,其中,核酸適配體8-3的親和力最好,其結構包含3個環狀部分和1個莖狀部分,其引物區域和環狀部分均為其與靶細胞結合的關鍵位點。此外,對其莖部進行適當截短后不影響其與靶細胞的結合能力,但是替換堿基后會使其失去對靶細胞的識別能力。結論:兔VX2細胞的核酸適配體有望應用于腫瘤發生發展機制、腫瘤示蹤以及靶向藥物遞送等方面的研究。
關鍵詞: 分析化學 核酸適配體 Cell-SELEX VX2細胞 結構優化 性能考察
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Selection and Structural Optimization of Aptamers against Rabbit VX2 cell
Abstract:Objective: to select the aptamers of rabbit VX2 cells, optimize the structure and investigate the characteristics of the obtained aptamers. Methods: using Cell-SELEX technology, rabbit VX2 cells were used as target cells and rabbit renal RK13 cells as control cells; After cutting and replacing the sequence of the obtained aptamer, its performance was investigated. Results: after 12 rounds of selecting, five aptamers of rabbit VX2 cells were obtained. Among them, the affinity of aptamer 8-3 was the best. Its structure contained three ring parts and one stem part. The primer region and ring part were the key sites for its binding with target cells. In addition, proper truncation of its stem does not affect its binding ability with target cells, but it will lose its recognition ability of target cells after base replacement. Conclusion: the aptamer of rabbit VX2 cells is expected to be used in the study of tumorigenesis and development mechanism, tumor tracing and targeted drug delivery.
Keywords: analytical chemistry aptamer Cell-SELEX rabbit VX2 cell structural optimization performance investigation
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兔VX2細胞的核酸適配體篩選和結構優化
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