Table 1.
Primer sequences used for single-guide RNA cloning, sequencing, and Surveyor assay
| Primer | Sequence (5′-3′) | Function |
|---|---|---|
| sgRNA2a for pX458 | CACCGTCATCCTCCTGACAATCGATAAACATCGATTGTCAGGAGGATGAC | Single-guide cloning |
| sgRNA1a for MulE vector | ACCGCAATGTGTCAACTCTTGACAAAACTGTCAAGAGTTGACACATTG | Single-guide cloning |
| sgRNA2a for MulE vector | ACCGTCATCCTCCTGACAATCGATAAACATCGATTGTCAGGAGGATGA | Single-guide cloning |
| hU6_Seq | GGACTATCATATGCTTACCGTAACTTGA | Primer for sequencing |
| CCR5-F | CCTGCCAAAAAATCAATGTGA | Surveyor assay/sequencing |
| CCR5-R | AGGACCAGCCCCAAGATGAC | Surveyor assay/sequencing |
| CCR2-F | GCAAATTGGGGCCCAACTCC | Surveyor assay/sequencing |
| CCR2-R | CCAAAATGTTCCTCATTATTGTGTGG | Surveyor assay/sequencing |
a
These oligos were annealed to make the double-stranded linker with 5′ overhangs (underlined nucleotides).