# Mobile-CRISPRi protocol optimization for Vibrionaceae

## TL;DR

This paper describes optimized methods for using the Mobile-CRISPRi system to knock down gene expression in various Vibrio bacteria.

## Contribution

The study provides optimized protocols for applying Mobile-CRISPRi in multiple Vibrio species.

## Key findings

- Mobile-CRISPRi components were successfully integrated into the chromosome of Vibrio species via Tn7 transposition.
- The optimized protocol enables inducible gene expression knockdown in multiple Vibrio species.
- The system uses isopropyl β-D-1-thiogalactopyranoside to induce deactivating Cas9 and guide RNA expression.

## Abstract

Mobile clustered regularly interspaced palindromic repeats interference (Mobile-CRISPRi) is an established method for bacterial gene expression knockdown. The deactivated Cas9 protein and guide RNA are isopropyl β-D-1-thiogalactopyranoside inducible, and all components are integrated into the chromosome via Tn7 transposition. Here, we optimized methods specific for applying Mobile-CRISPRi in multiple Vibrio species.