DNA-Free Genome Editing

Genetic engineering techniques, especially for CRISPR/Cas9 nuclease system, have greatly enhanced the ability of scientists to make genomic modifications. However, the public are often concerned about the safety of genetically modified (GM) food. Therefore, DNA-free genome editing methods have been developed and gained increasing attention in both academia and industrial field.

Lifeasible, as a leading plant biotechnology company, provides a variety of DNA-free genome editing methods to help you create safer and cleaner GMO products. These techniques include transiently expressed CRISPR/Cas9 plasmids DNA (TECCDNA) (Figure 1), CRISPR/Cas9 in vitro transcripts (IVTs), and preassembled ribonucleoprotein complexes (RNPs) consisting of purified Cas9 protein and transcriptional or synthetic sgRNAs (Figure 2). These methods can avoid foreign DNA integration into the host genomes and reduce off-target effects. Moreover, these techniques are cheaper and faster than conventional techniques by avoiding cross or backcross to segregate the CRISPR/Cas9 cassettes.

Figure 1. Overview of the genome-editing methods based on transiently expressing CRISPR/Cas9 DNA (Zhang et al., 2016).

Figure 2. DNA-free genome editing with CRISPR/Cas9 RNAs and RNP in plants (Yin et al., 2017).

We also provide many methods for the delivery of TECCDNA, IVTs, and/or RNPs into plants, including polyethylene glycol (PEG), cationic lipid, and/or electroporation mediated transfection of protoplasts and biolistic bombardment of immature embryos (Figure 1 and 2). Our team has used these techniques to transfect various types of plant tissues including protoplasts of Arabidopsis thaliana, tobacco, lettuce, and rice, as well as immature embryos and/or calli in wheat. With excellent experts, we can provide our customers worldwide with one-stop services from preparation and delivery of TECCDNA, IVTs, and RNPs to validation by PCR/restriction enzyme (RE) and next-generation sequencing. Our DNA-free genome editing services are proudly featured with high quality and efficiency. Please feel free to contact us for further information.