CRISPR-Cas9 Editing Induces Loss of Heterozygosity in the Pathogenic Yeast Candida parapsilosis.

Genetic manipulation is often used to study gene function. However, unplanned genome changes (including single nucleotide polymorphisms [SNPs], aneuploidy, and loss of heterozygosity [LOH]) can affect the phenotypic traits of the engineered strains. Here, we compared the effect of classical deletion methods (replacing target alleles with selectable markers by homologous recombination) with CRISPR-Cas9 editing in the diploid human-pathogenic yeast Candida parapsilosis.

Insights into the transcriptomic response of the plant engineering bacterium Ensifer adhaerens OV14 during transformation.

The ability to engineer plant genomes has been primarily driven by the soil bacterium Agrobacterium tumefaciens but recently the potential of alternative rhizobia such as Rhizobium etli and Ensifer adhaerens OV14, the latter of which supports Ensifer Mediated Transformation (EMT) has been reported. Surprisingly, a knowledge deficit exists in regards to understanding the whole genome processes underway in plant transforming bacteria, irrespective of the species. To begin to address the issue, we undertook a temporal RNAseq-based profiling study of E.

Ensifer-Mediated Transformation (EMT) of Rice (Monocot) and Oilseed Rape (Dicot).

Ensifer adhaerens OV14 underpins the successful crop transformation protocol, termed Ensifer-mediated transformation (EMT). The adaptability and efficiency of EMT technology to successfully transform both monocot and dicots have been previously reported. To facilitate community users' transition to EMT, the modified rice and oilseed rape plants generated in this work were developed using EMT protocols that were grounded in standard Agrobacterium-mediated transformation (AMT) processes.

Ensifer-mediated transformation: an efficient non-Agrobacterium protocol for the genetic modification of rice.

While Agrobacterium-mediated transformation (AMT) remains the most widely used technique for gene transfer in plants, interest exists for the use of non-Agrobacterium gene delivery systems due to freedom-to-operate issues that remain with AMT across several jurisdictions. In addition, the plant pathogenic mode of action of Agrobacterium tumefaciens significantly increases the costs to passage engineered cultivars through the regulatory process. Ensifer adhaerens (OV14) is a soil-related bacterium with the proven ability to genetically modify the model plant A.