Skip to content
DraconiticResearch
Search Sources API
203,203 studies31 sourcesUpdated 53 min ago
Peer-reviewedCC BYTrAC Trends in Analytical Chemistry

Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics

  • Wenyu Fu1
  • Rui Sang1
  • Ruier Xue1
  • Guochen Bao2
  • Shilun Feng3
  • Chunlei Jiao4,5,6,7,8
  • Fei Deng1
8 affiliations
  1. 1School of Biomedical Engineering, UNSW Sydney, Sydney 2052, Australia
  2. 2Institute for Biomedical Materials and Devices (IBMD), School of Mathematical and Physical Sciences, Faculty of Science, University of Technology Sydney, Sydney, New South Wales, Australia
  3. 3State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China
  4. 4Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117599
  5. 5Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, 117558
  6. 6NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), National University of Singapore, 117456, Singapore
  7. 7National Centre for Engineering Biology (NCEB), Singapore, 117575
  8. 8Synthetic Biology Translational Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117456
Open at sourceDOICite

Abstract

CRISPR-Cas systems have emerged as powerful biosensing platforms capable of highly sensitive and specific nucleic acid detection. The performance of these assays critically depends on the design of the guide RNA (gRNA), which governs target recognition and activates cis - and trans -cleavage. However, native gRNAs often exhibit limited stability, weak mismatch discrimination, reduced activity in clinical matrices, and inconsistent collateral cleavage, thereby limiting the diagnostic potential of CRISPR sensors. To address these constraints, a wide spectrum of gRNA engineering strategies has been developed, including length change, structural reconfiguration, DNA modified strategies, chemical modifications, mismatch as well as multiple modifications. These modifications substantially enhance sensitivity, specificity, stability, and programmability across Cas12a, Cas13a, and emerging CRISPR effectors. This review systematically summarizes recent advances, compares mechanistic principles and functional outcomes, and highlights future opportunities for rational gRNA engineering to enable amplification-free detection, improve clinical robustness, and drive the development of next-generation CRISPR diagnostics.

Related

  1. Dual‐Site Cu(II) Metalation in Donor−Acceptor Hydrogen‐Bonded Organic Framework Enables Record High‐Gain Photoelectrochemical Transistor for Ultrasensitive Biosensing
    Lu Hou, Chengshuang Wang, Xu Zhang +4
    Angewandte Chemie International EditionPeer-reviewedLicence not statedFull text7 citedMolecular biology & gene regulationOpenAlex
  2. Engineering a surfactant-free Au@nitrogen-doped graphene quantum dot heterointerface for ultrasensitive electrochemical detection of miR-9
    Nesa Rafati, Sahar Jafarzadeh, Emad Khademi +3
    RSC advancesPeer-reviewedCC BY-NCFull textMolecular biology & gene regulationEurope PMCEurope PMCPMC
  3. AI-integrated nanomaterial-based biosensors for biomedical diagnostics: engineering design, signal processing, and clinical translation.
    Chandrashekar Thalluri, Sarad Pawar Naik Bukke, Jharna Medhi +6
    Biomedical engineering onlinePeer-reviewedCC BY-NC-NDFull text2 citedMolecular biology & gene regulationEurope PMCDOAJ
  4. Engineering catalytic promiscuity in serum albumin: synergistic roles of microenvironment, structural dynamics, and nano conjugation.
    Swapan Patra, Nilanjan Dey
    RSC advancesPeer-reviewedCC BY-NCFull textMolecular biology & gene regulationEurope PMC
  5. Electrochemical in-biosensing computing
    Cheng Yuan, Ao Xiao, Shuang Wu +4
    National Science ReviewPeer-reviewedCC BY15 citedMolecular biology & gene regulationOpenAlex

Cite

BibTeXRIS
BibTeX
@article{fu2026engineering,
  title = {{Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics}},
  author = {Wenyu Fu and Rui Sang and Ruier Xue and Guochen Bao and Shilun Feng and Chunlei Jiao and Fei Deng},
  journal = {TrAC Trends in Analytical Chemistry},
  year = {2026},
  month = jun,
  doi = {10.1016/j.trac.2026.118960},
  url = {https://doi.org/10.1016/j.trac.2026.118960},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics
AU  - Wenyu Fu
AU  - Rui Sang
AU  - Ruier Xue
AU  - Guochen Bao
AU  - Shilun Feng
AU  - Chunlei Jiao
AU  - Fei Deng
PY  - 2026
DA  - 2026/06/05
JO  - TrAC Trends in Analytical Chemistry
DO  - 10.1016/j.trac.2026.118960
UR  - https://doi.org/10.1016/j.trac.2026.118960
AB  - CRISPR-Cas systems have emerged as powerful biosensing platforms capable of highly sensitive and specific nucleic acid detection. The performance of these assays critically depends on the design of the guide RNA (gRNA), which governs target recognition and activates cis - and trans -cleavage. However, native gRNAs often exhibit limited stability, weak mismatch discrimination, reduced activity in clinical matrices, and inconsistent collateral cleavage, thereby limiting the diagnostic potential of CRISPR sensors. To address these constraints, a wide spectrum of gRNA engineering strategies has been developed, including length change, structural reconfiguration, DNA modified strategies, chemical modifications, mismatch as well as multiple modifications. These modifications substantially enhance sensitivity, specificity, stability, and programmability across Cas12a, Cas13a, and emerging CRISPR effectors. This review systematically summarizes recent advances, compares mechanistic principles and functional outcomes, and highlights future opportunities for rational gRNA engineering to enable amplification-free detection, improve clinical robustness, and drive the development of next-generation CRISPR diagnostics.
SN  - 0165-9936
SN  - 0167-2940
SN  - 1879-3142
ER  -