AMPK Methods and Protocols

PRKAA1: chromosome 5p

3’UTR

Exon 7

PRKAA1exon 7

TTCAACCATGATTGATGATGAAGCCTTAAAAGAAGTATGTGAAAAGTTTGAGTGCTCAGAAG

AGGAAGTTCTCAGCTGTCTTTACAACAGAAATCACCAGGATCCTTTGGCAGTTGCCTACCA

TCTCATAATAGATAACAGGAGAATAATGAATGAAGCCAAAGATTTCTATTTGGCGACAAGCCC

ACCTGATTCTTTTCTTGATGATCATCACCTGACTCGGCCCCATCCTGAAAGAGTACCATTCT

TGGTTGCTGAAACACCAAGGGCACGCCATACCCTTGATGAATTAAATCCACAGAAATCCAA

ACACCAAGGTGTAAGGAAAGCAAAATGGCATTTAGGAATTAGAAGTCAAAGTCGACCAAAT

GATATTATGGCAGAAGTATGTAGAGCAATCAAACAATTGGATTATGAATGGAAG

AMPKα1 sgRNA#1

AMPKα1 sgRNA#2

PRKAA1 foward sequencing primer

PRKAA1 reverse sequencing primer

Underlined: 239 bp predicted amplicon.

A

B

PRKAA2 exon 1

GCGCCCAGCGCCCTCTCCCGGGACCCCGCGCCCGCCCCGCGCCCCTCGCGGCTCGGGG

TCCGCCCCCTCGCCCGCAGGGCCCGCTGCACTGTGGGTAGGCGGCGGCGGCGGCGGCT

ACGCGGAGCGGCAGGCGGTGGAGCGAGGCCGCGCGCGCCGAAGATGGCTGAGAAGCA

GAAGCACGACGGGCGGGTGAAGATCGGACACTACGTGCTGGGCGACACGCTGGGCGTC

GGCACCTTCGGCAAAGTGAAGAGTTGAGTACGCGCTCCGGACCGCTGTGCGGGGCTGCC

TGACTTGCGGGAGGCCGAGGGGAGAGGCGGGAGCCCCGGGCCTCCGGCGGGCGCGGG

AMPKα2 sgRNA#1

AMPKα2 sgRNA#2

PRKAA2 foward sequencing primer

PRKAA2 reverse sequencing primer

Underlined: 158 bp predicted amplicon.

PRKAA2: chromosome 5p

3’UTR

Exon 1

Fig. 1Graphical representation ofPRKAA1andPRKAA2genomic DNA sequences.PRKAA1andPRKAA2
encompass 9 and 10 exons, respectively. Exons are figured asvertical black rectangles.PRKAA1 exon 7 is
highlighted inred. Intronic regions are depicted by connecting roof-shaped lines. 3^0 untranslated regions
(3^0 UTR) are indicated. The scale is indicated, 1kd: 10 bases (A) AMPKα1 sgRNA#1 (green) and sgRNA#2 (red),
forward (blue) and reverse (pink) sequencing primers, and predicted amplicon (underlined) are represented.
(B) AMPKα2 sgRNA 1 (green) and 2 (red), forward (blue) and reverse (pink) sequencing primers, and predicted
amplicon (underlined) are represented

CRISPR-Cas9 For Human AMPK 177