RAPID-DASH: fast and efficient assembly of guide RNA arrays for multiplexed CRISPR-Cas9 applications

TL;DR

This paper introduces a fast method to build gRNA arrays for CRISPR-Cas9, enabling efficient targeting of multiple genes in a single day.

Contribution

A novel, rapid method for constructing scalable and functional gRNA arrays with up to 10 units.

Findings

gRNA arrays maintain functional activity across all positions.

The method allows for the incorporation of gRNA libraries, enabling scalable and multiplexed targeting.

A web tool is provided to design oligo sequences for gRNA array assembly.

Abstract

Guide RNA (gRNA) arrays can enable targeting multiple genomic loci simultaneously using CRISPR-Cas9. In this study, we present a streamlined and efficient method to rapidly construct gRNA arrays with up to 10 gRNA units in a single day. We demonstrate that gRNA arrays maintain robust functional activity across all positions, and can incorporate libraries of gRNAs, combining scalability and multiplexing. Our approach will streamline combinatorial perturbation research by enabling the economical and rapid construction, testing, and iteration of gRNA arrays. To facilitate the adoption of this approach, we have made a web tool to design oligo sequences necessary to assemble gRNA arrays.

Graphical Abstract

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