Table 4 Oligonucleotides used in this study
From: CRISPR/Cas9 Knockout Strategies to Ablate CCAT1 lncRNA Gene in Cancer Cells
| Gene | Sequence (5′ to 3′) | Product (bP) | Application |
|---|---|---|---|
| CCAT1 (NR_108049.1) |
F: CTGACAACATCGACTTTG R: CTCACAGTTTTCAAGGGA Probe: FAM-CTTAGCCATACAGAGCCAACCTG-BHQ1 | 108 | qPCR |
|
F: CGATCGttctgttttcaatggggatt R: TCGAGggagctgcggataacagcatat | 546 | Cloning of left homology arm | |
|
F: CTAGTCCCgcatcacagctactgtcaaccc R: ATCCCCtcaaagcacttctgtggtagga | 832 | Cloning of right homology arm | |
|
Forward-1: CACATGGCTCCCATCACACTA Reverse-1: GGGGGAAGAAATTTAAGATGCACA | 1298 (w.t.) 409 (ko) | PCR confirmation of CRISPR Excision knockout allele | |
|
Forward-2: CACGCAGATCACATGACCCT Reverse-2: CGGGCCATTTACCGTAAG | 1627 (ko) | PCR confirmation of CRISPR HDR knockout allele | |
|
Forward-2: CACGCAGATCACATGACCCT R: AAACgatggagctgcggataacagC | 843 (w.t.) | PCR confirmation of CRISPR HDR wild-type allele | |
|
Forward-3: CGGGCCATTTACCGTAAG Reverse-1: GGGGGAAGAAATTTAAGATGCACA | 495 (ko) | PCR confirmation of CRISPR du-HITI knockout allele | |
|
F: CACCGaatcggagtccaaagccatt R: AAACaatggctttggactccgattC | – | sgRNA (downstream of Exon 1) | |
|
F: CACCGataatggaggggatttacgt R: AAACacgtaaatcccctccattatC | – | sgRNA (upstream of Exon 1) | |
|
F: CACCGctgttatccgcagctccatc R: AAACgatggagctgcggataacagC | – | sgRNA (Exon 2) | |
|
F: CACCGaatggctttggactccgatttgg R: AAACccaaatcggagtccaaagccattC | – | sgRNA bait (+PAM) (downstream of Exon 1) | |
| b-Actin |
F: TGCAGAAGGAGATCACTG R: CTTGCTGATCCACATCTG Probe: CY5-AAGATCAAGATCATTGCTCCTCCTGA-BHQ2 | 141 | qPCR |