cDNA primers (RT-qPCR) | |||||
---|---|---|---|---|---|
Gene | Species | Forward sequence | Reverse sequence | Template length | Notes |
Gapdh mRNA | H. Sapiens | TGTCAAGCTCATTTCCTGGTAT | CTCTCTTCCTCTTGTGCTCTTG | 133 bp | Intron- spanning |
GRM2 mRNA | H. Sapiens | CATTCCTGCCCATCTTCTATGT | GTGGCTAACCACGTTCTTCT | 161 bp | Intron- spanning |
Gapdh mRNA | R. norvegicus | ACCTTTGATGCTGGGGCTGGC | GGGCTGAGTTGGGATGGGGACT | 217 bp | Intron- spanning |
Tent5b mRNA | R. norvegicus | CCAGCAGATTGTACAGGTAGT | CTCACTCTGCAGGTCCATCT | 163 bp | Intron- spanning |
Gadd45b mRNA | R. norvegicus | GACAACGCGGTTCAGAAGAT | TCCTCCTCTTCTTCGTCTATGG | 173 bp | Intron- spanning |
Sstr2 mRNA | R. norvegicus | CGTGGAAAAGCAAGATGTCAC | TATGGCTCTGTCTGGTTGGA | 213 bp | Intron- spanning |
gDNA primers | |||||
Gene | Location | Forward sequence | Reverse sequence | Template length | |
Human GAPDH | 642 bp upstream of GAPDH TSS | GAGCCTCGAGGAGAAGTTCC | GGACCCTTACACGCTTGGAT | 167 bp | |
Rat Gapdh | 174 bp upstream of Gapdh TSS | GGTCGGAGCCCACACGCTTG | TCCCGCTCGGCTCATCCAGT | 161 bp | |
“P1” inverted DIO-dCas9 | Forward primer in beginning of dCas9 with reverse primer in WRPE | GGCCGATGCTGTACTTCTTG | TGTAATCCAGAGGTTGATTGTC | 324 bp | |
“P2” correct orientation, recombined DIO-dCas9 | Forward primer in hSYN with reverse primer in beginnig of dCas9 | GAGTCGTGTCGTGCC | GGCCGATGCTGTACTTCTTG | 180 bp (constitutive), 242-bp DIO | |
“P3” inverted SVI-DIO | Forward primer in hSYN with reverse primer in inverted first segment of dCas9 | GAGTCGTGTCGTGCC | ATTCTGCGGCGGCAGGAAGA | 359 bp | |
“P4” recombined SVI-DIO | Intron-spanning in recombined SVI-DIO | ACCTTCCGCATCCCCTACTA | TCGAAGTTGGTCATCCGCTCG | 262 bp (spliced)/ 421 bp (unspliced) | |
CRISPR guide RNA (sgRNA) | |||||
Genomic locus | Species | Target sequence | Genomic coordinates | ||
lacZ | E. coli | TGCGAATACGCCCACGCGAT | E. coli strain 99–3165:3,886, 546–3,886,565 | ||
Tent5b | R. norvegicus | GAATTCGGTCCGAAGAGAAG | Rn6: Chr5: 155,354,031 | ||
Gadd45b | R. norvegicus | GGCGGTGGAGCAGCTGCTGG | Rn6: Chr7: 11,647,911–11,647,930 | ||
Sstr2 | R. norvegicus | TGCGCCGCTAGCATTGGCCG | Rn6: Chr10: 102,136,282–102,136,263 | ||
Fos | R. norvegicus | GAGCGGAACAGAGAAACTGG | Rn6: Chr6: 109,300,373–109,300,354 | ||
Fos | R. norvegicus | GTGAAAGTTACAGACTGAGA | Rn6: Chr6: 109,300,259–109,300,278 | ||
Fos | R. norvegicus | GTGCAGCGCAAGGGGGGAGC | Rn6: Chr6: 109,300,064–109,300,045 | ||
Fos | R. norvegicus | GCCTAATGTTGCACTGATTT | Rn6: Chr6: 109,299,775–109,299,794 |