CRISPR-Cas系統新的應用:SHERLOCK檢測技術
華裔科學家張鋒利用CRISPR-Cas新系統,研發出了新的快速,便宜,高靈敏度的診斷工具,可以用于檢測諸如寨卡病毒感染和登革熱感染等的疾病,這一進展也有助于便捷檢測和診斷許多其它的病原體,該結果發表在著名雜志科學(Science)上。不同于Cas9,新的Cas蛋白是Cas13a,具有在剪切掉目標RNA后還能保持活性,可以“插手”剪切其他非目標RNA的特點。運用Cas13a的“連帶剪切”特性,再添加一種特定RNA熒光標記物,并使用特殊擴增技術,依靠體溫擴增檢測樣本中的RNA,就形成了一個準確檢測到單個核酸分子的檢測系統。
該系統命名為“神探夏洛克(SHERLOCK,Specific High Sensitivity Enzymatic Reporter UnLOCKing)”??梢杂糜跈z測血清、尿液和唾液中低濃度的寨卡病毒,而且可以用于檢測病毒載量,區分不同的菌株,甚至能分辨獨有不同耐藥突變的肺炎菌株和不同的癌癥突變。
又是一篇科學(Science)文章,祝賀華裔大神在“神”的道路上繼續高速前進。
原文摘要:
Nucleic acid detection with CRISPR-Cas13a/C2c2
Abstract:Rapid, inexpensive, and sensitive nucleic acid detection may aid point-of-care pathogen detection, genotyping, and disease monitoring. The RNA-guided, RNA-targeting CRISPR effector Cas13a (previously known as C2c2) exhibits a "collateral effect" of promiscuous RNAse activity upon target recognition. We combine the collateral effect of Cas13a with isothermal amplification to establish a CRISPR-based diagnostic (CRISPR-Dx), providing rapid DNA or RNA detection with attomolar sensitivity and single-base mismatch specificity. We use this Cas13a-based molecular detection platform, termed SHERLOCK (Specific High Sensitivity Enzymatic Reporter UnLOCKing), to detect specific strains of Zika and Dengue virus, distinguish pathogenic bacteria, genotype human DNA, and identify cell-free tumor DNA mutations. Furthermore, SHERLOCK reaction reagents can be lyophilized for cold-chain independence and long-term storage, and readily reconstituted on paper for field applications.