Plant Multiplex Gene Editing Service

We are committed to becoming your reliable assistant and partner in the field of plant protein

  1. Home
  2. Services
  3. Plant Genetic Engineering Services
  4. Plant Gene Editing Services
  5. Plant Multiplex Gene Editing Service
Services
Online Inquiry

Plant Multiplex Gene Editing Service

Inquiry

The development of multiple genome editing systems is essential for the stacking or improvement of agronomic traits in plants and for the control of gene regulatory pathways. CD BioSciences has mature CRISPR gene editing systems. Our CRISPR Cas12a platform can simultaneously target multiple genes at specific locations in plants. For large-scale genome engineering, we also provide clients with Cas12a system development and optimization services to improve gene editing efficiency.

Figure 1. Modeling of the CRISPR-Cas12a catalytic pathway. – CD BioSciences  Figure 1. Modeling of the CRISPR-Cas12a catalytic pathway. (Paul, B., & Montoya, G., 2020)

Service Content

We offer the Cas12a system with a potential target site next to the PAM (5'-TTV-3', V for A/C/G) sequence, which once encountered with Cas12, creates base pair hybridization between the crRNA and the target DNA strand to form an R-loop. Then, Cas12a utilizes the RuvC domain to cleave the non-target strand. In addition, Cas12a's crRNA is significantly shorter than Cas9's sgRNA, allowing for the formation of compact Cas12a CRISPR arrays that can be used to target multiple loci simultaneously.

Cas12a vs Cas9

Type Cas12a Cas9
Size of protein 1,100~1,300 amino acids 1,000~1,600 amino acids
Nuclease sites Single nuclease site RuvC-Nuc Two nuclease domains HNH and RuvC
RNA crRNA crRNA and tracrRNA
Type of cut Staggered ends Blunt ends
PAM requirements Recognizes 5' T-rich PAM sequences of 3-4 nt Recognizes 3' G-rich PAM sequences of 3-5 nt
precrRNA processing Possesses intrinsic RNase activity to process precr-RNA Requires host RNase III and tracrRNA

Experiment procedure

  • Design crRNA so that its spacer is complementary to the DNA target sequence and custom synthesize crRNA.
  • Mix crRNA with Cas12a protein to form RNP.
  • Introduce RNP into the plant cell or nucleus.
  • Recognize the DNA target sequence by the spacer of the crRNA and the PAM sequence by the Cas12a protein to shear the DNA.
  • Repair of DNA breaks.
    1) Perform non-homologous un-terminal junction repair to achieve gene knockout.
    2) Designing donor DNA templates and performing homologous recombination repair to achieve gene knock-in, knock-out, silencing, etc.

Advantages of the Cas12a System

  • The CRISPR-Cas12a system expands the editing site and generates interleaved cuts at the 5' end of the sequence, significantly increasing the probability that the cell will choose homologous recombination repair (HDR) and enhancing the efficiency of gene editing.
  • Cas12a can perform genome editing in organisms with an AT-rich genome with a wider range of sequence editing.
  • Only a shorter crRNA is required to complex with Cas12a without tracrRNA, which is more favorable for chemical synthesis.

As a leading biotech company in the gene editing industry, CD BioSciences has accumulated rich experience in the field of plant gene editing. With our cutting-edge equipment and proven CRISPR gene editing systems, we can guarantee the success of clients' projects. If you have requirements in plant gene modification, please contact us for more details.

References

  1. Paul, B., & Montoya, G. (2020). CRISPR-Cas12a: Functional overview and applications. Biomedical Journal.43(1),8-17.
  2. Bandyopadhyay, A., et al. (2020). CRISPR-Cas12a (Cpf1): A Versatile Tool in the Plant Genome Editing Tool Box for Agricultural Advancement. Frontiers in plant science. 11, 584151.

For research use only, not for clinical use.