# RAG recombinase expression discriminates the development of natural killer cells

## TL;DR

This study shows that RAG recombinase activity, previously thought irrelevant to natural killer cells, actually plays a role in their development and function.

## Contribution

The study demonstrates that RAG expression contributes to the maturation of natural killer cells, challenging prior assumptions.

## Key findings

- GFP expression in mature NK cells indicates RAG activity during their development.
- Transcriptomic analysis reveals advanced maturation and compromised DNA damage response in RAG-expressing NK cells.
- RAG-expressing NK cells show strong effector function but reduced survival after ionizing radiation.

## Abstract

V(D)J recombination, initiated by recombination-activating gene (RAG) endonucleases, is a crucial process for the generation of diversified antigen receptors of T and B lymphocytes but regarded dispensable for innate natural killer (NK) lymphocytes lacking clonotypic receptors.

To explore the impact of potential rearrangements on NK cell maturation, RAG-fate mapping reporter human induced pluripotent stem cell (iPSC) lines were generated by introduction of RSS-invEGFP constructs into the AAVS1 locus using CRISPR/Cas9 and differentiated into NK cells in vitro.

GFP expression was observed in up to 14% of mature NK cells characterized by a CD45dim CD56dimCD57+NKG2C+/−KIR+/− phenotype and unproductive genetic rearrangements in the IGH locus. Advanced maturation was further revealed by transcriptomic studies using RNA sequencing. Despite their strong effector function, DNA damage response and survival to ionizing radiation were compromised.

These findings suggest a role of RAG expression in NK cell ontogeny supporting the development of a terminally differentiated effector population.