Limited budget, no other constraints |
Two-part guide RNA |
High endo- or exo-nuclease activity in cells |
Single guide RNA (first choice) |
OR (next choice if the first choice does not work), two-part guide RNA with many chemical modifications,* |
OR (third choice, if the above choices do not work), single guide RNA with many chemical
modifications** |
Delivery of pre-formed Cas9 protein |
Two-part guide RNA, OR single guide RNA (both choices are equally likely to work) |
Delivery of mRNA or plasmid coding for Cas9 |
Single guide RNA for longer stability |
Low editing efficiency with single guide RNA |
Two-part guide RNA (first choice), |
OR try another target site (second choice if the first choice does not work) |
Low editing efficiency with two-part guide RNA |
Single guide RNA (first choice), |
OR try another target site (second choice if the first choice does not work) |