Efficient targeted DNA editing and replacement in Chlamydomonas reinhardtii using Cpf1 ribonucleoproteins and single-stranded DNA

Significance Our findings establish a method of efficient, targeted genome editing in Chlamydomonas reinhardtii . We demonstrate an approach to bypass inefficient gene targeting via homologous recombination and achieve homology-directed DNA replacement in C. reinhardtii . In addition, we report CRISPR/Cpf1-mediated DNA editing efficiencies being boosted 500-fold through the use of single-stranded oligodeoxynucleotides (ssODNs) as repair templates. It remains to be determined whether Cpf1-induced staggered DNA cleavage enhances ssODN-mediated gene editing in a wider range of species and whether the underlying repair pathway(s) responsible is more broadly conserved.