Journal
PLANTS-BASEL
Volume 10, Issue 10, Pages -Publisher
MDPI
DOI: 10.3390/plants10102055
Keywords
CRISPR interference; crop improvement; gene silencing; RNAi; transcriptional regulation; CRISPR/dCas9 system
Categories
Funding
- Ministry of Higher Education Malaysia [FRGS/1/2018/STG03/UM/02/2, FP065-2018A]
- Universiti Malaya [ST003-2021, RU005-2019, ST009-2019]
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RNA-guided genomic transcriptional regulation tools, CRISPRi and CRISPRa, derived from the CRISPR/Cas9 system, utilize a catalytically dead Cas9 to achieve transcriptional interference. Despite its potential benefits, challenges and limitations such as off-target effects, PAM sequence requirements, and labeling as genetically modified organisms are faced by the CRISPR/dCas9 system.
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.
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