| RRC ID |
89422
|
| Author |
Oishi S, Doi S, Sekida T, Yamashita K, Yamada Y, Muramoto T.
|
| Title |
Genome editing across Dictyostelia species enables comparative functional genetics of social amoebas.
|
| Journal |
Sci Rep
|
| Abstract |
Gene manipulation is essential for understanding biological mechanisms, yet genetic modification in the social amoebas (Dictyostelia) has been largely limited to Dictyostelium discoideum. Here, we aimed to establish a CRISPR/Cas9-based genome-editing system applicable across the phylogenetic breadth of Dictyostelia, spanning Groups 1-4. Using an extrachromosomal CRISPR/Cas9 vector from D. discoideum, we disrupted stlA and pkaC in Polysphondylium violaceum and pkaC in two early-branching species, Heterostelium pallidum and Cavenderia fasciculata. In D. discoideum, co-introduction of donor oligos with the CRISPR vector enabled selection-free knockout generation of pkaC with 28.6% efficiency. In H. pallidum, where genome editing is typically inefficient, co-electroporation of donor oligos with the CRISPR/Cas9 vector followed by 4 days of drug selection increased the frequency of pkaC disruption from 0.9% to 8.3%. These results demonstrated that the D. discoideum CRISPR/Cas9 system can be extended across Dictyostelia, providing a versatile platform for comparative genetic and evolutionary developmental studies.
|
| Volume |
16(1)
|
| Published |
2026-2-5
|
| DOI |
10.1038/s41598-026-38605-5
|
| PII |
10.1038/s41598-026-38605-5
|
| PMID |
41644992
|
| PMC |
PMC12929561
|
| MeSH |
CRISPR-Cas Systems*
Dictyostelium* / genetics
Gene Editing*
Genome, Protozoan*
Phylogeny
|
| Resource |
| Cellular slime molds |
|
