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Updated: May 31, 2026
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
Compact Cas12f enables genome editing in avian cells
Yujin Han1, Seung Je Woo1, Hee Jung Choi1
1Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, the Republic of Korea.
Poultry Science
|May 29, 2026
Summary
The compact CRISPR Cas12f nuclease demonstrates efficient genome editing in avian cells, overcoming delivery challenges of larger systems like Cas9. This tool shows high on-target efficiency with minimal off-target effects, paving the way for avian genetic engineering.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- CRISPR genome editing in avian species is limited by the large size and low delivery efficiency of nucleases like Cas9 and Cas12a.
- The compact Cas12f nuclease offers enhanced delivery efficiency in mammalian systems, but its avian application was unproven.
Purpose of the Study:
- To evaluate the effectiveness and safety of the compact Cas12f nuclease for genome editing in avian cells.
- To assess Cas12f's transfection efficiency, cytotoxicity, on-target editing, indel profiles, and off-target mutations in chicken cells.
Main Methods:
- Cas12f was delivered into chicken Leghorn male hepatoma (LMH) cells and primordial germ cells (PGCs).
- Transfection efficiency, intracellular expression, and cytotoxicity were assessed.
- Next-generation sequencing (NGS) and Sanger sequencing with Inference of CRISPR Edits (ICE) were used for on-target and off-target analysis.
Main Results:
- Cas12f exhibited notable transfection efficiency and intracellular expression in LMH cells and PGCs without detectable cytotoxicity.
- On-target editing efficiencies reached up to 40% in a locus-dependent manner in LMH cells.
- Cas12f generated a deletion-dominant indel profile, distinct from Cas9, with minimal off-target mutations detected above the threshold.
Conclusions:
- Cas12f is a viable and efficient compact genome editing tool for avian cells.
- Its high delivery efficiency and specificity make it a promising alternative to larger CRISPR nucleases for avian genetic engineering and biotechnology.
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