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Catalytic Hairpin Assembly-Assisted Rolling Circle Amplification for High-Sensitive Telomerase Activity Detection.
ACS Omega ( IF 3.7 ) Pub Date : 2020-05-12 , DOI: 10.1021/acsomega.0c01459 Yang Liu 1 , Shihong Li 1 , Likun Zhang 1 , Qian Zhao 1 , Nuo Li 1 , Yuxin Wu 1
ACS Omega ( IF 3.7 ) Pub Date : 2020-05-12 , DOI: 10.1021/acsomega.0c01459 Yang Liu 1 , Shihong Li 1 , Likun Zhang 1 , Qian Zhao 1 , Nuo Li 1 , Yuxin Wu 1
Affiliation
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Telomerase is a promising biomarker and a potential therapeutic target of malignant tumors. Reliable, facile, and sensitive telomerase activity analysis is of vital importance for both early diagnosis and therapy of malignant tumors. Herein, we proposed a novel fluorescent assay termed catalytic hairpin assembly-assisted rolling circle amplification (CAR) for both in vitro and in situ high-sensitive telomerase activity detection. In the presence of active telomerase, the extension of a designed telomerase primer was limited to five bases (GGGTT), thus forming short telomerase products. Afterward, the obtained telomerase extension products cyclized Padlock and subsequently initiated the rolling circle amplification (RCA). In order to maintain a higher sensitivity, an ingeniously designed catalytic hairpin assembly (CHA) was attached for both signal amplification and result readout. The highlights of the CAR method were concluded as follows: (i) dual signal amplification from CHA and RCA ensures high sensitivity and (ii) the CAR method has the potential for both in vitro and intracellular imaging of telomerase activity. We believe that the CAR method would be of great potential for the diagnosis and therapy of various human diseases.
中文翻译：
用于高灵敏度端粒酶活性检测的催化发夹组装辅助滚环扩增。
端粒酶是一种有前途的生物标志物，是恶性肿瘤的潜在治疗靶标。可靠，简便，敏感的端粒酶活性分析对于恶性肿瘤的早期诊断和治疗均至关重要。在这里，我们提出了一种新型的荧光测定法，称为催化发夹装配辅助滚环扩增（CAR），用于体外和原位高灵敏度端粒酶活性检测。在存在活性端粒酶的情况下，设计的端粒酶引物的延伸限于五个碱基（GGGTT），从而形成短的端粒酶产物。之后，获得的端粒酶延伸产物使挂锁环化，随后启动滚环扩增（RCA）。为了保持较高的灵敏度，一个巧妙设计的催化发夹组件（CHA）被连接用于信号放大和结果读出。CAR方法的亮点归纳如下：（i）CHA和RCA的双重信号扩增可确保高灵敏度，并且（ii）CAR方法具有对端粒酶活性进行体外和细胞内成像的潜力。我们相信，CAR方法对于各种人类疾病的诊断和治疗具有巨大的潜力。
更新日期：2020-05-12
中文翻译：
用于高灵敏度端粒酶活性检测的催化发夹组装辅助滚环扩增。
端粒酶是一种有前途的生物标志物，是恶性肿瘤的潜在治疗靶标。可靠，简便，敏感的端粒酶活性分析对于恶性肿瘤的早期诊断和治疗均至关重要。在这里，我们提出了一种新型的荧光测定法，称为催化发夹装配辅助滚环扩增（CAR），用于体外和原位高灵敏度端粒酶活性检测。在存在活性端粒酶的情况下，设计的端粒酶引物的延伸限于五个碱基（GGGTT），从而形成短的端粒酶产物。之后，获得的端粒酶延伸产物使挂锁环化，随后启动滚环扩增（RCA）。为了保持较高的灵敏度，一个巧妙设计的催化发夹组件（CHA）被连接用于信号放大和结果读出。CAR方法的亮点归纳如下：（i）CHA和RCA的双重信号扩增可确保高灵敏度，并且（ii）CAR方法具有对端粒酶活性进行体外和细胞内成像的潜力。我们相信，CAR方法对于各种人类疾病的诊断和治疗具有巨大的潜力。
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