Engineering a CRISPR-associated IscB system for developing miniature genome-editing tools in human cells and mouse embryos

Fei Zhang (), Yuxin Peng, Dongdong Fan, Guoxu Song, Xing Gao and Yong Tian ()
Additional contact information
Fei Zhang: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics
Yuxin Peng: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics
Dongdong Fan: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics
Guoxu Song: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics
Xing Gao: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics
Yong Tian: Chinese Academy of Sciences, State Key Laboratory of Epigenetic Regulation and Intervention, Institute of Biophysics

Nature Communications, 2025, vol. 16, issue 1, 1-16

Abstract: Abstract IscB, as the putative ancestor of Cas9, possesses a compact size, making it suitable for in vivo delivery. OgeuIscB is the first IscB protein known to function in eukaryotic cells but requires a complex TAM (NWRRNA). Here, we characterize a CRISPR-associated IscB system, named DelIscB, which recognizes a flexible TAM (NAC). Through systematically engineering its protein and sgRNA, we obtain enDelIscB with an average 48.9-fold increase in activity. By fusing enDelIscB with T5 exonuclease (T5E), we find that enDelIscB-T5E displays robust efficiency comparable to that of enIscB-T5E in human cells. Moreover, by fusing cytosine or adenosine deaminase with enDelIscB nickase, we establish efficient miniature base editors (ICBE and IABE). Finally, we efficiently generate mouse models by microinjecting mRNA/sgRNA of enDelIscB and enDelIscB-T5E into mouse embryos. Collectively, our work presents a set of enDelIscB-based miniature genome-editing tools with great potential for diverse applications in vivo.

Date: 2025
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-025-65724-w Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-65724-w

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-025-65724-w

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().