Multiplexed CRISPR technologies for gene editing and transcriptional regulation.

Nature Communications
Nicholas S McCartyRodrigo Ledesma-Amaro

Abstract

Multiplexed CRISPR technologies, in which numerous gRNAs or Cas enzymes are expressed at once, have facilitated powerful biological engineering applications, vastly enhancing the scope and efficiencies of genetic editing and transcriptional regulation. In this review, we discuss multiplexed CRISPR technologies and describe methods for the assembly, expression and processing of synthetic guide RNA arrays in vivo. Applications that benefit from multiplexed CRISPR technologies, including cellular recorders, genetic circuits, biosensors, combinatorial genetic perturbations, large-scale genome engineering and the rewiring of metabolic pathways, are highlighted. We also offer a glimpse of emerging challenges and emphasize experimental considerations for future studies.

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Methods Mentioned

BETA
transfect
PCRs
PCR
to
produce
GUIDE-seq
biosensors
biosensing
RNA-seq

Software Mentioned

Benchling
BLESS
WeReview CRISPR Tools
CAMERA
CHOPCHOP
GUIDE
Geneious
Digenome

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