Compact RNA-Guided Nucleases

This repository contains reference constructs for compact and ultra-compact RNA-guided nucleases for in vivo genome editing. Examples highlight well-established reference designs that are commonly used as baselines or starting points for optimization.

Staphylococcus aureus Cas9 (SaCas9)

• Native SaCas9 nuclease Wild-type bacterial coding sequence derived from the ORFX locus of Staphylococcus aureus.

• Human-codon–optimized SaCas9 nuclease Standalone coding sequence derived from the SaCas9 used in early in vivo editing studies.

• AAV SaCas9 reference design Single-vector AAV expression cassette derived from the architecture reported by Ran et al. (Nature, 2015). Provided as a historical and practical baseline illustrating payload constraints for Cas9-based in vivo editing.

CasMINI (engineered Cas12f)

• CasMINI expression construct (CAG promoter, nuclease + gRNA) Single-plasmid mammalian expression design adapted from peer-reviewed validation studies. This construct demonstrates a functional ultra-compact RNA-guided nuclease system in mammalian cells.

Constructs in this repository are provided as reusable design primitives. Where applicable, sequences have been curated to remove target-specific elements or reporters. Additional compact nucleases, engineered variants, and delivery-oriented designs (e.g., AAV-compatible CasMINI systems) will be added.