Table 1.
sgRNA No. | Exon No. | Strand | sgRNA Target Sequences, Excluding PAM (5′–3′) | sgRNA Target Sequence Position | PAM Nucleotide Position | Cut Sites in the DMD Gene | Cut Sites in the Amino Acid Sequence | sgRNA Used in the Formation of the Hybrid Exon |
---|---|---|---|---|---|---|---|---|
1 | 46 | sense | TTCTCCAGGCTAGAAGAACAA | 1407207–1407227 | 1407228–1407233 | 6624 – 6225 | 2208 GAA (Glu): 2209 CAA (Gln) | 46–51; 46–53 |
2 | 46 | antisense | CTGCTCTTTTCCAGGTTCAAG | 1407312–1407332 | 1407306–1407311 | 6714 –6715 | 2238 CTT (Leu): 2239 GAA (Glu) | 46–53 |
3 | 47 | sense | GTCTGTTTCAGTTACTGGTGG | 1409686–1409706 | 1409707–1409712 | 6769 –6770 | 2257 G:TG (Val) | 47–58 |
4 | 47 | antisense | TCCAGTTTCATTTAATTGTTT | 1409736–1409756 | 1409730–1409735 | 6824 –6825 | 2268 AAA (Lys): 2267 CAA (Gln) | 47–58 |
5 | 47 | antisense | CTTATGGGAGCACTTACAAGC | 1409765–1409785 | 1409759–1409764 | 6833 – 6834 | 2278 CT:T (Leu) | 47–58 |
6 | 49 | antisense | TTGCTTCATTACCTTCACTGG | 1502716–1502736 | 1502710–1502715 | 7194 –7195 | 2398 CCA (Pro): 2399 GTG (Val) | 49–52; 49–53 |
7 | 51 | antisense | TTGTGTCACCAGAGTAACAGT | 1565282–1565302 | 1565276–1564281 | 7323 –7324 | 2441 ACT (Thr): 2442 GTT (Val) | 46–51 |
8 | 51 | antisense | AGTAACCACAGGTTGTGTCAC | 1565294–1565314 | 1565288-1565293 | 7335 –7336 | 2445 GTG (Val): 2446 ACA (Thr) | 46–51 |
9 | 52 | antisense | TTCAAATTTTGGGCAGCGGTA | 1609765–1609785 | 1609759–1609764 | 7595 –7596 | 2532 AC:C (Thr) | 47–52 |
10 | 52 | sense | CAAGAGGCTAGAACAATCATT | 1609802–1609822 | 1609823–1609828 | 7647 −7648 | 2549 ATC (Ile): 2550 ATT (Ile) | 49–52 |
11 | 53 | antisense | TTGTACTTCATCCCACTGATT | 1659891–1659911 | 1659885–1659890 | 7677 – 7678 | 2559 AAT (Asn): 2560 CAG (Gln) | 49–53 |
12 | 53 | sense | CTTCAGAACCGGAGGCAACAG | 1659918–1659938 | 1659939–1659944 | 7719 – 7720 | 2573 CAA (Gln): 2574 CAG (Gln) | 46–53 |
13 | 53 | sense | CAACAGTTGAATGAAATGTTA | 1659933–1659953 | 1659954–1659959 | 7734 –7735 | 2578 ATG (Met): 2579 TTA(Leu) | 46–53 |
14 | 53 | sense | GCCAAGCTTGAGTCATGGAAG | 1660017–1660037 | 1660038–1660043 | 7818 –7819 | 2606 TGG (Trp): 2607 AAG (Lys) | 46–53 |
15 | 53 | antisense | CTTGGTTTCTGTGATTTTCTT | 1660068–1660088 | 1660062–1660067 | 7854 – 7855 | 2618 AAG (Lys): 2619 AAA (Lys) | 46–53 |
16 | 58 | sense | TCATTTCACAGGCCTTCAAGA | 1860349–1860369 | 1860370–1860375 | 8554 –8555 | 2852 A:AG (Lys) | 47–58 |
17 | 58 | antisense | CAGAAATATTCGTACAGTCTC | 1860411–1860431 | 1860405–1860410 | 8601 –8602 | 2867 GAG (Gln): 2868 ACT (Thr) | 47–58 |
18 | 58 | antisense | CAATTACCTCTGGGCTCCTGG | 1860467–1860487 | 1860461–1860466 | 8657 – 8658 | 2886 GT:C (Gln) | 47–58 |
Nucleotide positions are provided with reference to the DMD gene sequence ENS00000198947 analyzed using the Benchling web tool (https://benchling.com/). Bold underlined sequences are intron sequences.