Associate Professor Yizhen Liu and Professor Xueji Zhang Publish Important CRISPR Diagnosis Achievements in Nature Biomedical Engineering-College of Chemistry and Envionmental Engineering

Associate Professor Yizhen Liu and Professor Xueji Zhang Publish Important CRISPR Diagnosis Achievements in Nature Biomedical Engineering

Author: Date: 2025-07-01 15:32 click: [ ]

Recently, Associate Professor Yizhen Liu and Professor Xueji Zhang from the College of Chemistry and Environmental Engineering have made breakthroughs in the field of CRISPR gene diagnosis technology, providing innovative technical solutions for clinical precision diagnosis. The related achievements, titled "LbuCas13a directly targets DNA and elicits strong trans-cleavage activity", were published in the top international academic journal Nature Biomedical Engineering (Impact Factor: 26.6). Xiaolong Wu and Siyuan Luo, master's students in Chemistry (2022 cohort) supervised by Associate Professor Yizhen Liu, are the first authors. Associate Researcher Yong Chen, Associate Professor Yizhen Liu, and Professor Xueji Zhang serve as corresponding authors, with the College of Chemistry and Environmental Engineering at Shenzhen University as the sole authoring unit.

link: https://www.nature.com/articles/s41551-025-01424-6

Traditional theory holds that Cas13a, as an RNA-specific nuclease, can only recognize RNA targets. The research team discovered that the LbuCas13a protein derived from Leptotrichia buccalis can directly bind to DNA targets and generate strong trans-cleavage activity, endowing the detection method with excellent sensitivity. Moreover, it does not rely on the protospacer adjacent motif (PAM) or flanking sequences (PFS), breaking through the sequence limitations of traditional CRISPR technology. More importantly, when LbuCas13a binds to single-stranded DNA, the reduced affinity between guide RNA (crRNA) and DNA enhances its single-nucleotide specificity by 98-fold compared to RNA targeting. The discovery of these characteristics makes LbuCas13a an excellent tool for precise genotyping, laying an important foundation for the development of accurate diagnostic technologies for major diseases.

Based on the above findings, the team developed the "Superior Universal Rapid Enhanced Specificity Test" platform (SUREST, LbuCas13a-based Superior Universal Rapid Enhanced Specificity Test). This technology exhibits extremely high resolution for single-nucleotide polymorphism (SNP) detection. Accurate SNP genotyping is crucial for personalized medicine. Clinical sample testing shows that for genotyping multiple human genes related to drug metabolism and disease susceptibility, such as CYP1A1 and CYP2C9, as well as highly specific identification of viral mutation sites (K417N, S477N, T478K, Q498R, N501Y, Y505H mutations, etc.), the genotyping results of the SUREST platform are completely consistent with the gold standard Sanger sequencing. The DNA-targeting property of LbuCas13a without PAM sequence restrictions enables CRISPR diagnosis to truly achieve highly specific genotyping for any target, fundamentally expanding the clinical application boundaries of gene detection.

The SUREST technology integrates recombinase polymerase amplification (RPA) with the strong trans-cleavage activity of LbuCas13a to achieve extremely high detection sensitivity. The sensitivity for detecting the CYP2C19 gene (rs4986893) reaches 0.3 aM (0.18 copies/μL), and it can complete the quantitative analysis of hepatitis B virus and HPV16 virus within more than 20 minutes, with a detection limit as low as 1 aM.

The SUREST platform demonstrates high sensitivity, high specificity, and full target coverage in clinical testing, showing clear application value in precision medicine scenarios such as rapid genotyping of pathogen strains, precise identification of rare disease-causing mutations, drug metabolism gene-guided medication, and ultra-early cancer screening. It is expected to promote the popularization of precision medical analysis technologies in primary healthcare settings and facilitate the early diagnosis and treatment of major diseases.

This research was supported by the National Key Research and Development Program, Shenzhen Medical Research Special Fund Project, National Natural Science Foundation of China, Guangdong Basic and Applied Basic Research Foundation, and Shenzhen Key Laboratory of Nanobiosensing Technology. The clinical samples for this study were provided by the Third People's Hospital of Shenzhen and South China Hospital Affiliated to Shenzhen University.