Application of CRISPR-Cas9 System in the Plant Biology Field

The CRISPR-Cas9 genome editing system consists of a “guide” RNA (gRNA) and a non-specific CRISPR-associated endonuclease (Cas9). The Cas9 protein is an endonuclease that uses gRNA to form base pairs with DNA target sequences, enabling Cas9 to introduce a site-specific double-stranded break (DSB) in the DNA. The CRISPR-Cas9 system can modify DNA with greater precision than existing technologies like TALEN and ZFN.

The CRISPR-Cas9 system can be used to increase the productivity of plantations and protect natural forests from further deforestation. Traditional conifer breeding methods are time-consuming due to the long juvenile period and complex genome. While the CRISPR/Cas9 system provides a way to rapidly alter key traits in conifers through genome editing. Using plasmid-expressed Cas9 gRNA and RNPs, respectively, can generate biallelic and monoallelic INDELs in conifer P. radiata.RNPs are co-delivered with plasmid-based selectable markers. Using a dual gRNA strategy, an editing efficiency of 33% was achieved and INDELs containing large deletions were generated.

Figure 1. Editing of GUX1 with a single guide RNA.

Figure 2. Editing of GUX1 with two guide RNAs.

Figure 3. Details of editing in GUX1 using RNPs.

In recent years, the CRISPR-Cas9 system has attracted extensive attention due to its simple operation and high efficiency, along with its wide applications in the regulation of gene mutation and transcriptional regulation in mammals and plants. Therefore, CRISPR/Cas is considered a revolutionary technology.

CRISPR-Cas9 Gene Editing Services | Synbio Technologies

Synbio Technologies, as a synthetic biology enabled company, provides one-stop CRISPR-Cas9 gene editing services, including CRISPR sgRNA design, oligo synthesis, sgRNA library construction, NGS verification, and bioinformatics analysis. At present, Synbio Technologies has delivered hundreds of high-quality CRISPR sgRNA libraries covering dozens of species including animals, plants, and microorganisms. According to the different needs of customers, the customized libraries can be knockout libraries, interference libraries, or activation libraries, committing to better serve the customer’s downstream researches.

Reference
Charleson Poovaiah, Lorelle Pillips, Barbara Geddes, et al. Genome editing with CRISPR/Cas9 in Pinus radiata (D. Don). BMC Plant Biol. 2021; 21: 363. DOI: 10.1186/s12870-021-03143-x