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Updated: May 3, 2026

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
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DNA-guided CRISPR-Cas12a effectors for programmable RNA recognition and cleavage.

Xiaolong Wu1, Wai Hei Lam2, Zibin Zhao1

  • 1Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, China.

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|May 1, 2026
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Summary
This summary is machine-generated.

Researchers engineered CRISPR-Cas12a to target RNA using DNA guides. This novel DNA-guided RNA targeting system enables direct RNA detection and cellular RNA knockdown, expanding CRISPR applications.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • CRISPR-Cas systems commonly use RNA guides for target recognition.
  • Cas12a activation involves DNA protospacer adjacent motif interaction.
  • Existing systems primarily target DNA or RNA using RNA guides.

Purpose of the Study:

  • To reprogram CRISPR-Cas12a into a DNA-guided, RNA-targeting effector.
  • To investigate the molecular mechanisms of this novel targeting configuration.
  • To establish a new platform for programmable RNA manipulation.

Main Methods:

  • Engineering synthetic CRISPR DNA for Cas12a complex formation.
  • Utilizing structural, biophysical, and biochemical analyses.
  • Assessing intracellular RNA detection and knockdown efficiency.

Main Results:

  • Successfully reprogrammed Cas12a to function as a DNA-guided, RNA-targeting system.
  • Elucidated the molecular basis of DNA-guided RNA targeting and its distinct activation pathway.
  • Demonstrated efficient direct RNA detection and intracellular RNA knockdown using the engineered system.

Conclusions:

  • The engineered Cas12a expands the design space for programmable RNA manipulation.
  • This work establishes a modular activation architecture for CRISPR-Cas12a.
  • DNA-guided RNA targeting offers new possibilities for molecular diagnostics and therapeutics.