Evaluation of Alcalase pretreatment for Chlamydomonas reinhardtii CRISPR knock-in
We wanted to use CRISPR-Cas9-mediated homology-directed repair (HDR) in C. reinhardtii to insert genomic landing pads as part of a larger project. In cell-walled algae, delivering Cas9 ribonucleoproteins (RNPs) and donor DNA requires first digesting that wall. The standard approach uses autolysin, a gamete-derived prot...