Skip to content
DraconiticResearch
Search Sources API
203,203 studies31 sourcesUpdated 14 min ago
Peer-reviewedCC BY-NC-NDBiodesign research

Precision meets scale: DSB- and recombinase-free targeted insertion of large DNA fragments

  • Koppolu Raja Rajesh Kumar1
  • Ayushi Singh1
  1. 1Department of Biotechnology, Indira Gandhi National Tribal University, Amarkantak, India.
Read full textPDFDOICite

Abstract

Targeted insertion of large DNA fragments has been constrained by the genotoxicity associated with double-strand breaks (DSBs) and the limitations of two-step recombinase systems. Three independently developed mammalian platforms, Quadruple pegRNA (QuadPE), Prime Assembly (PA), and PRIME-In, now achieve programmable, DSB- and recombinase-free insertion of up to 26 kb, 11.3 kb, and 9.2 kb, respectively, through convergent yet mechanistically distinct flap-guided integration strategies. Notably, QuadPE and PA achieve integration independent of cell-cycle progression. Furthermore, these systems outperform several existing recombinase- and/or transposase-based tools, underscoring their translational potential for therapeutic utility. Their mechanistic logic also provides a forward path for the DSB-free and recombinase-free integration of large DNA sequences in crop biotechnology.

Related

  1. Targeted insertion of partial CFTR cDNA replicates native CFTR expression with restored channel function in CF airway epithelium
    Cristina Barillà, Shingo Suzuki, So Hyun Park +20
    Molecular therapy. Nucleic acidsPeer-reviewedCC BYFull textMolecular biology & gene regulationEurope PMC
  2. Targeted insertion of an optimized donor DNA is effective in a humanized mouse model of dominant retinitis pigmentosa
    Federica Esposito, Arjun Padmanabhan, Manuel Rhiel +14
    Cell Reports MedicinePeer-reviewedCC BY-NC-ND 4.0Full textMolecular biology & gene regulationPMC
  3. Targeted insertion of partial
    Cristina Barillà, Shingo Suzuki, So Hyun Park +20
    Molecular Therapy: Nucleic AcidsPeer-reviewedCC BYFull textMolecular biology & gene regulationPharmacology & toxicologyDOAJEurope PMCPMC
  4. Targeted Modulation of the Gut Microbiota in Obesity: From Dietary Interventions to Bacteriophage-Based Precision Therapies
    Sthefhany Nohemí Rodríguez-Arellano, Jean Pierre González-Gómez, José Roberto Aguirre-Sánchez +4
    NutrientsPeer-reviewedCC BY 4.0Molecular biology & gene regulationCrossref
  5. Characterization of recombinase-based genetic parts and circuits using nanopore sequencing.
    F Veronica Greco, Sarah K Cameron, Shivang Hina-Nilesh Joshi +4
    Nature communicationsPeer-reviewedCC BYFull text2 citedMolecular biology & gene regulationEurope PMC

Cite

BibTeXRIS
BibTeX
@article{kumar2026precision,
  title = {{Precision meets scale: DSB- and recombinase-free targeted insertion of large DNA fragments}},
  author = {Koppolu Raja Rajesh Kumar and Ayushi Singh},
  journal = {Biodesign research},
  year = {2026},
  month = oct,
  doi = {10.1016/j.bidere.2026.100126},
  url = {https://europepmc.org/article/PMC/PMC13718414},
  note = {CC BY-NC-ND},
}
RIS
TY  - JOUR
TI  - Precision meets scale: DSB- and recombinase-free targeted insertion of large DNA fragments
AU  - Koppolu Raja Rajesh Kumar
AU  - Ayushi Singh
PY  - 2026
DA  - 2026/10/10
JO  - Biodesign research
DO  - 10.1016/j.bidere.2026.100126
UR  - https://europepmc.org/article/PMC/PMC13718414
AB  - Targeted insertion of large DNA fragments has been constrained by the genotoxicity associated with double-strand breaks (DSBs) and the limitations of two-step recombinase systems. Three independently developed mammalian platforms, Quadruple pegRNA (QuadPE), Prime Assembly (PA), and PRIME-In, now achieve programmable, DSB- and recombinase-free insertion of up to 26 kb, 11.3 kb, and 9.2 kb, respectively, through convergent yet mechanistically distinct flap-guided integration strategies. Notably, QuadPE and PA achieve integration independent of cell-cycle progression. Furthermore, these systems outperform several existing recombinase- and/or transposase-based tools, underscoring their translational potential for therapeutic utility. Their mechanistic logic also provides a forward path for the DSB-free and recombinase-free integration of large DNA sequences in crop biotechnology.
SN  - 2693-1257
ER  -