Table 1.
List of gRNA sequences and Cas9 plasmids used for core-clock (CC) KO in HCT116 cells. For each target gene, multiple gRNAs binding different genomic regions were used to increase KO efficiency.
| Target Gene | gRNA Seq (without PAM)—(5′ –> 3′) | Cas9 Type |
|---|---|---|
| ARNTL | ACAGACAAAGATGACCCTCA | pSpCas9(BB)-2A-GFP |
| ARNTL | TTATCACACTACGGAGTCGA | pSpCas9(BB)-2A-GFP |
| ARNTL | CTGGACATTGCGTTGCATGT TAGATAAACTTACTGTGCTA |
Cas9D10A-GFP (AIO-GFP) |
| PER2 | GACCAACGAAAACTGCTCCA | pSpCas9(BB)-2A-GFP |
| PER2 | GAACACAACCCATCTACAAG | pSpCas9(BB)-2A-GFP |
| PER2 | CCCCGTGGAGCAGTTTTCGT GCAGTGACTGTGACGACAGT |
Cas9D10A-GFP (AIO-GFP) |
| NR1D1 | GTTGCGATTGATGCGGACGA | pSpCas9(BB)-2A-GFP |
| NR1D1 | CGTAGGTGAAGATCTCTCGA | pSpCas9(BB)-2A-GFP |