Spatiotemporal regulation of clustered regularly interspaced short palindromic repeats (CRISPR) system is attractive for precise gene editing and accurate molecular diagnosis. Although many efforts have been made, versatile and efficient strategies to control CRISPR system are still desirable. Here, we proposed a universal and accessible acylation strategy to regulate the CRISPR–Cas12a system by efficient acylation of 2′-hydroxyls (2′-OH) on crRNA strand with photolabile agents (PLGs). The introduction of PLGs confers efficient suppression of crRNA function and rapid restoration of CRISPR–Cas12a reaction upon short light exposure regardless of crRNA sequences. Based on this strategy, we constructed a universal PhotO-Initiated CRISPR–Cas12a system for Robust One-pot Testing (POIROT) platform integrated with recombinase polymerase amplification (RPA), which showed two orders of magnitude more sensitive than the conventional one-step assay and comparable to the two-step assay. For clinical sample testing, POIROT achieved high-efficiency detection performance comparable to the gold-standard quantitative PCR (qPCR) in sensitivity and specificity, but faster than the qPCR method. Overall, we believe the proposed strategy will promote the development of many other universal photo-controlled CRISPR technologies for one-pot assay, and even expand applications in the fields of controllable CRISPR-based genomic editing, disease therapy, and cell imaging.
The aim of this randomized controlled study was to investigate the effects of a novel external catheter fixation method for chemotherapy using inferior epigastric arterial catheterization for cervical cancer.Patients diagnosed with cervical cancer were randomly divided into a control group (n = 32) and a treatment group (n = 33). Patients in the control group underwent a traditional fixation method using a haemostat, elastic band and abdominal bandage. Patients in the treatment group underwent an improved fixation method using an indwelling needle and membrane cover. We used a visual analogue scale (VAS) to evaluate each patient's comfort score and also recorded the incidence of needlestick injury and the length of injection time in each group. The VAS scores measured before and after chemotherapy in the treatment group were lower than in the control group. The incidence of needlestick injury in the treatment group was significantly lower than in the control group. The length of injection time in treatment group was significantly lower than in the control group. Compared with the traditional fixation method, the improved fixation method not only increased patient comfort but also reduced both the risk of needlestick injury and the length of injection time. This improved technique deserves increased clinical use.