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Review
. 2021 Sep 29;10(10):2055.
doi: 10.3390/plants10102055.

CRISPR/dCas9-Based Systems: Mechanisms and Applications in Plant Sciences

Chou Khai Soong Karlson  1 , Siti Nurfadhlina Mohd-Noor  2 , Nadja Nolte  3 , Boon Chin Tan  1
Affiliations
Review

CRISPR/dCas9-Based Systems: Mechanisms and Applications in Plant Sciences

Chou Khai Soong Karlson et al. Plants (Basel). .

Abstract

RNA-guided genomic transcriptional regulation tools, namely clustered regularly interspaced short palindromic repeats interference (CRISPRi) and CRISPR-mediated gene activation (CRISPRa), are a powerful technology for gene functional studies. Deriving from the CRISPR/Cas9 system, both systems consist of a catalytically dead Cas9 (dCas9), a transcriptional effector and a single guide RNA (sgRNA). This type of dCas9 is incapable to cleave DNA but retains its ability to specifically bind to DNA. The binding of the dCas9/sgRNA complex to a target gene results in transcriptional interference. The CRISPR/dCas9 system has been explored as a tool for transcriptional modulation and genome imaging. Despite its potential applications and benefits, the challenges and limitations faced by the CRISPR/dCas9 system include the off-target effects, protospacer adjacent motif (PAM) sequence requirements, efficient delivery methods and the CRISPR/dCas9-interfered crops being labeled as genetically modified organisms in several countries. This review highlights the progression of CRISPR/dCas9 technology as well as its applications and potential challenges in crop improvement.

Keywords: CRISPR interference; CRISPR/dCas9 system; RNAi; crop improvement; gene silencing; transcriptional regulation.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic illustration showing sgRNA, dCas9 and transcriptional effectors. dCas9 together with sgRNA forms the dCas9/sgRNA complex for transcriptional regulator attachment. Fusion of the complex with effectors, transcription activators or repressors, are used for targeted gene regulation.
Figure 2
Figure 2
Schematic diagram showing the difference between the CRISPR/Cas9 system and the CRISPR/dCas9 system utilizing Cas9 and deactivated Cas9 (dCas9), respectively.
Figure 3
Figure 3
A schematic diagram shows the multiplexability of the CRISPR/dCas9 system. Enhancement of transcriptional regulation, simultaneous targeting of different genes and simultaneous targeted binding at multiple loci are possible using multiple sgRNAs.
Figure 4
Figure 4
Different types of dCas9 protein with or without the effectors, such as KRAB and VP64.
See this image and copyright information in PMC

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