Table 1.
List of primer sequences
gRNA competition assay | |
---|---|
Human gRNAs | Sequence |
SRPK1 (1) | GGTGTGGATGATACGGCACT |
SRPK1 (2) | CTGCATGGTATTTGAAGTTT |
SRPK1 (3) | TTACCGGTCTCACCATGGAG |
BRD4S-ex12-SA_A | TTTCTCTCTCCCTCTACGT |
BRD4S-ex12-SA_B | TTAGGCAGGACCTACGTAG |
BRD4_ET_domain | AGTCGATTTCAATCTCGTCG |
BRD4_BD2_domain | GTAGAAGGGCCAGGCGTAGG |
BCL2 Intergenic (1) | GAGTGTCTCAATGGGCAGCG |
BCL2 Intergenic (2) | AAGAGCCACGGCCTAAAGCA |
Mouse gRNAs | Sequence |
Srpk1 (1) | ACCTGCAGACCCCGATGGTG |
Srpk1 (2) | TGAATGAGCAGTACATTCGA |
RT-PCR | ||
---|---|---|
Forward Primer | Reverse Primer | |
HOXA9 | GAATGAGAGCGGCGGAGAC | GAGCGAGCATGTAGCCAGTTG |
c-MYC | AATGAAAAGGCCCCCAAGGTAGTTATCC | GTCGTTTCCGCAACAAGTCCTCTTC |
BCL2 | CTGCACCTGACGCCCTTCACC | CACATGACCCCACCGAACTCAAAGA |
BRD4 | CTCCTCCTAAAAAGACGAAGA | TTCGGAGTCTTCGCTGTCAGAGGAG |
GAPDH | GATGCCCTGGAGGAAGTGCT | AGCAGGCACAACACCACGTT |
RIP qPCR | ||
---|---|---|
Forward Primer | Reverse Primer | |
BRD1 | ACATCCCATGGACTTTGCCACA | CTCACCGCGGCTCTATAGAACA |
BRD4 | CTCCTCCTAAAAAGACGAAGA | TTCGGAGTCTTCGCTGTCAGAGGAG |
MYB | AGCACCGATGGCAGAAAGTACT | TCTCCCCTTTAAGTGCTTGGCA |
MED24 | AGGAGCTCAAGTGGACAGCTTT | GGTGAGCTTCAGCAGGAACTCA |
ChIP qPCR | ||
---|---|---|
Forward Primer | Reverse Primer | |
BCL2 | ACAGCGCCAACAGAACTACT | CCCCACAACGGAGCTGTAAT |
MYC | CAAGCTCTCCACTTGCCCCT | GCCCTGAGATGTGTCTGCCT |
EZH2 | CTTCTGAGTCCCACCGGGTG | GCCGTGTGTTCAGCGAAAGA |
DUSP6 | GTAGAGGAAGGTCGGGGAGA | CACACAGGGCCATCTCAACT |
DGKI | GCCACCCCCTCATCTCTCAC | TCTTCCAAGGACCCAGGGGA |
Splicing Validation – RT-qPCR | ||
---|---|---|
Exon Skipping | Forward Primer | Reverse Primer |
BRD1 (Exons 11-12) | AATGTCACTGAGGTCGCTGG | ACGTGCTGAAGATTGGGGAG |
BRD4-Long (Exons 10-12) | TCGGAGCCATCTCTGTTTC | CGACTTTGAGACCCTGAAGC |
BRD4-Short (Exons 10-12) | AATGATTAGGCAGGACCTGTT | CGACTTTGAGACCCTGAAGC |
Exon Inclusion | Forward Primer | Reverse Primer |
MYB (Exon 8-Intron 8) | TGAGCTAAAAGGACAGCAGGT | CAAAGCACAAGGAGCCATC |
Alternative 5' Splice Site | Forward Primer | Reverse Primer |
PIM1 (Exon 4-Intron 4) | CGACATCAAGGACGAAAACA | AGACACCCACACCCTTTCCT |
Intron Retention | Forward Primer | Reverse Primer |
MED24 (Exon 6-Intron 6) | AAGTCCCTGGGATGTGTGC | CTGGAGAAAACCCTCAGCAG |
KAT5 (Exon 1- Intron 1) | CGGAACCAGGACAACGAAGA | ACCTCTCGGAGCAGCTAAGA |
METTL3 (Exon 9-Intron 8) | AGACCCTGGTTGAAGCCTTG | TGGGGCCCAATTCAATAGGT |
VEGFA (Exon 5-Intron 5) | ACCAAAGAAAGATAGAGCAAGACAA | ATTGTTGCTGCCACCACAAG |
MED16 (Exon 14-Intron 14) | GACTTGAGCATGGTGACACAG | AAAGTGGGGCAGGCGTAA |
PRMT5 (Exon 12-Intron 11) | CACCCATTCCCTCATGTCTG | TTTCCTTAACATCTCTCCTTACCTT |