Efficient CRISPR-based method for targeted disruption of genes in Chlamydomonas
In May 2020, the Witman and Oda labs published a paper titled “TIM, a targeted insertional mutagenesis method utilizing CRISPR/Cas9 in Chlamydomonas reinhardtii” (PMC7219734). The paper describes an easy, rapid, and cost-effective method for efficiently targeting specific genes for disruption in Chlamydomonas. The authors tested TIM on six different genes in two different cell-walled strains and in each case disrupted the targeted gene with mutagenesis efficiencies ranging from 40% to greater than 90%. These very high efficiencies even enabled two different genes to be targeted in a single experiment. They expect the method will work on most non-essential nuclear genes.