Sugano, S. S., et al. · 2018 · PLoS ONE research
doi:10.1371/journal.pone.0205117 PMID:30379827 PMC6209168
| Gene ID | Name | Evidence / Role | Function (this paper) |
|---|---|---|---|
| Mp1g12750 | MpARF1 | experimental subject | Primary target for evaluating CRISPR/Cas9 editing efficiency; Mparf1 mutants show NAA-resistant phenotype enabling positive selection. Co-transformation yielded hundreds of NAA-resistant T1 plants, all 16 sequenced harbored indels/substitutions, with no off-target mutations at three predicted sites. |
| Mp2g08950 | MpMPK1 | experimental subject | Essential MAPK gene (knockouts unobtainable by gene targeting) studied via conditional knockout; CRISPR alone gave only mosaic mutants while co-transformation with cDNA complementation vectors yielded monoclonal frameshift mutants. Heat-shock/dexamethasone-induced Cre/loxP excision of the rescuing cDNA caused severe growth defects, confirming a critical role in growth and development. |
| Mp8g11800 | MpNOP1 | experimental subject | Targeted to assess editing efficiency via the easily scored transparent-thallus (air-chamber-less) phenotype; 87.5% of 32 T1 plants were mutated. Used to test guide length (17nt or fewer dramatically less efficient) and to induce 0.5/1.5/4.5kb deletions with paired gRNAs; no off-target mutations. |
| Mp3g23400 | MpEF1 | experimental tool | MpEF promoter drives Atco-Cas9 expression in editing vectors; preferentially expressed in meristematic tissues, immunoblot showed higher Cas9 accumulation than hCas9. |
| — | MpHSP17.8A1 missing | experimental tool | MpHSP17.8A1 heat-shock promoter drives Cre-GR for heat-shock/dexamethasone-inducible conditional recombination in the knockout system. |
| Mp5g03810 | MpPHOT | experimental tool | Mpphot (phototropin) detected by antibody as loading control in immunoblot analysis of Cas9 expression. |
| — | MpU6-1 missing | experimental tool | MpU6-1 promoter (2kbp and 500bp versions) drives gRNA expression; both lengths gave comparable genome-editing efficiency. |