Nucleic acids research

Using CRISPR-related transposons to design viral delivery tools and precise gene editing

Updated

Abstract

SHOT 2.0 achieves integration of large DNA cargos (at least 14 kb) into defined loci.

  • The optimized platform enables customizable bacmid editing in Escherichia coli.
  • Integration at the ODVe56 locus significantly enhances transgene stability during serial virus passaging.
  • The system allows for dual-gene insertion and is compatible with the Bac-to-Bac® workflow.
  • An all-in-one baculovirus was constructed to encode the PE5max prime editor.
  • efficiencies reached up to 85.6% in HEK293T cells and 37.1% in hard-to-transfect cell types.

Simplified

Key numbers

85.6%
Efficiency in HEK293T Cells
Efficiency of achieved using the SHOT 2.0 platform.
37.1%
Efficiency in Liver Cancer Cells
Efficiency achieved in hard-to-transfect liver cancer cells using the SHOT 2.0 platform.
14 kb
Large DNA Cargo Integration
Minimum size of DNA fragments that can be integrated using SHOT 2.0.

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