RNA-based CRISPR-CAS9 gene editing is a vector-free approach that is required for therapeutic perspectives. However, from a practical point of view, what appears as a major challenge to engineer primary cells, is actually also an efficient and smart solution for numerous other knock-out projects on cell lines. We see here why and how.

The idea to use RNA oligos as a therapeutic has been around since the late 1990s, however there is now an explosion of renewed interest. The Wall Street Journal‘s exclusive billion dollar club includes the RNA therapy companies Moderna and CureVac along side the likes of Uber and Airbnb.

Many labs have adopted the CRISPR genome editing technology to make knock-out and knock-in cell lines.

This technology produces first a targeted break in genomic DNA, which can then be exploited to produce cell lines with genes knocked out or where a donor vector has been used to introduce new genetic elements (point mutants, fluorescent tags, antibiotic resistance cassettes, etc.). Essentially any desired modification to the cells genome can be made. In setting up these genome editing projects there are many choices to be made including vector for the Cas9 protein and for the sgRNAs. Perhaps the most difficult choice, however, can be which cell line to use. Even the most affordable stable genome editing cell line development services can come with a significant cost, so choosing the right cell line at the beginning is crucial. Here we explain some of the choices researchers have in setting up their CRISPR genome editing projects and give our advice for cell line selection.

Many researchers are facing a dilemma: they want to set up a CRISPR genome editing project but they can’t decide which cell line to use for genome editing. Even some of the most cost-effective genome editing cell line generation services like the one from GeneCopoeia will cost a few thousand euros, so picking the correct cell line and setting up the project correctly is very important. Researchers working in primary cells may find the idea of switching to an immortalized cell line a bit artificial. They dream of the possibility of editing the genome of a stem cell line which they can then differentiate into their tissue of choice as needed.