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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
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Split CRISPR/Cas systems: Pioneering solutions for molecular diagnostics challenges.

Junqi Zhang1, Mengjia Zhu2, Hongbin Yan3

  • 1School of Life Sciences, State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei University, Wuhan, 430042, Hubei, China; Pilot Base of Food Microbial Resources Utilization of Hubei Province, School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, 430023, Hubei, China.

Biosensors & Bioelectronics
|October 31, 2025
PubMed
Summary
This summary is machine-generated.

Split CRISPR/Cas systems offer enhanced sensitivity and specificity for detecting low-abundance biomarkers without pre-amplification. These advanced molecular diagnostic tools show promise for precision diagnostics, especially in resource-limited settings.

Keywords:
CRISPR/Cas systemsMolecular diagnosticsSplit activatorSplit crRNA

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • Conventional CRISPR detection methods face limitations in sensitivity, specificity, and regulatory flexibility for low-abundance targets.
  • Split CRISPR/Cas systems represent an advancement to overcome these challenges in molecular diagnostics.

Purpose of the Study:

  • To review split-activation strategies in CRISPR/Cas systems for enhanced diagnostic performance.
  • To highlight innovations in ultrasensitive RNA detection and multiplex nucleic acid analysis.

Main Methods:

  • Focuses on split-activation strategies, including split activator-mediated Cas systems and split crRNA architectures.
  • Examines applications in clinical and on-site monitoring.

Main Results:

  • Achieves femtomolar-level sensitivity for biomarkers with superior single-base discrimination.
  • Enhances analytical performance without requiring pre-amplification.

Conclusions:

  • Split CRISPR/Cas systems offer significant improvements for precision diagnostics.
  • Future directions involve customized Cas variants, nanomaterial-based workflows, and microfluidic platforms for broader applications, especially in resource-limited settings.