chlamydomonas reinhardtii snapgene files

Previously, a null mutation of CpFTSY in C.

We first generated a specific knockout of the CpFTSY gene that confers a smaller, or truncated, chlorophyll (Chl) antenna size of the photosystems. Targeted CpFTSY gene knockout using RGEN RNPs reinhardtii, which has a fully sequenced and annotated genome and possesses haploid chromosomes 14, by using DNA-free RGEN RNPs. Here, we demonstrate targeted gene editing in C. In addition, the resulting gene-edited plants and animals could be exempt from genetically modified organism (GMO) regulations due to the absence of foreign DNA sequences 13. Furthermore, RGEN RNPs reduce off-target effects and mosaicism and may also be less cytotoxic in cells because the Cas9 protein is transiently active and then degraded by endogenous proteases in cells 9. The RGEN RNP delivery obviates the need for codon optimization or specific promoters in different species so that it can be conveniently and rapidly applied to various organisms. elegans 8, human cells 9, mice 10 and plants 11, 12.

Recently, DNA-free RGENs, that is, preassembled Cas9 protein-gRNA ribonucleoproteins (RNPs), have enabled efficient genome editing in C. However, although CRISPR RGENs are rapidly superseding ZFNs and TALENs due to the ease of application and low cost 7, a recent study with them showed that stable expression of Cas9 in the model green microalga Chlamydomonas reinhardtii is cytotoxic, resulting in targeted mutagenesis at a frequency below 10 −9 6.

For gene editing at desired target loci, programmable nucleases, such as zinc finger nucleases (ZFNs) 3, transcription-activator-like effector nucleases (TALENs) 4, 5 and RNA-guided engineered nucleases (RGENs) 6 derived from the type II CRISPR-Cas9 system have been applied to microalgae. Thus, genetic manipulations of microalgae for increasing a feedstock of bioenergy, natural resources of biomedical and biochemical compounds generation highly attract attention from both academic and industrial establishments. Unicellular microalgae have enormous potential as cell factories for the generation of biomass, fuel-type molecules and synthetic chemistry feedstock while, at the same time, helping to mitigate CO 2 emissions in the atmosphere 1, 2.

Chlamydomonas reinhardtii snapgene files