Networking to Optimize Dmd exon 53 Skipping in the Brain of mdx52 Mouse Model
Abstract
:1. Introduction
2. Materials and Methods
3. Results
3.1. In Vitro Screening of ASO Sequences Targeting Mouse Dmd Exon 53
3.2. Intramuscular Injection of Single or Combined ASO Targeting Exon 53 in mdx52 Mice
3.3. Intracerebroventricular Injection of Combined ASOs Targeting Exon 53 in mdx52 Mice
3.4. Combination of four ASOs Targeting Exon 53 Skipping Achieves Higher Level of Exon Skipping but Still Only Very Low Levels of Protein Restoration
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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CHEMISTRY | REGION | NAME | TARGET POSITION | ABBREVIATED NAME | TARGET SEQUENCE |
---|---|---|---|---|---|
tcDNA | PRE-SCREENING | tcDNA +27+41 | +27+41 | GAGGTTCAAGAACAG | |
tcDNA +33+47 | +33+47 | CAAGAACAGCTGCAG | |||
tcDNA +36+50 | +36+50 | GAACAGCTGCAGAAC | |||
tcDNA +38+52 | +38+52 | ACAGCTGCAGAACAG | |||
tcDNA +40+54 | +40+54 | AGCTGCAGAACAGGA | |||
tcDNA +41+55 | +41+55 | GCTGCAGAACAGGAG | |||
tcDNA +42+56 | +42+56 | CTGCAGAACAGGAGA | |||
tcDNA +43+57 | +43+57 | TGCAGAACAGGAGAC | |||
tcDNA +44+58 | +44+58 | GCAGAACAGGAGACA | |||
tcDNA +46+60 | +46+60 | AGAACAGGAGACAAC | |||
tcDNA +59+73 | +59+73 | ACAGTTGAATGAAAT | |||
tcDNA +73+87 | +73+87 | TGTTAAAGGATTCAA | |||
R1 | tcDNA +33+52 | +33+52 | CAAGAACAGCTGCAGAACAG | ||
tcDNA +36+55 | +36+55 | tcDNA_R1a | GAACAGCTGCAGAACAGGAG | ||
tcDNA +39+58 | +39+58 | CAGCTGCAGAACAGGAGACA | |||
tcDNA +42+61 | +42+61 | CTGCAGAACAGGAGACAACA | |||
tcDNA +45+64 | +45+64 | tcDNA_R1b | CAGAACAGGAGACAACAGTT | ||
tcDNA +56+75 | +56+75 | ACAACAGTTGAATGAAATGT | |||
R2 | tcDNA +69+88 | +69+88 | GAAATGTTAAAGGATTCAAC | ||
tcDNA +72+91 | +72+91 | tcDNA_R2 | ATGTTAAAGGATTCAACACA | ||
tcDNA +75+94 | +75+94 | TTAAAGGATTCAACACAATG | |||
tcDNA +78+97 | +78+97 | AAGGATTCAACACAATGGCT | |||
tcDNA +100+120 | +100+120 | AAGCTAAGGAAGAAGCCGAA | |||
R3 | tcNDA +125+144 | +125+144 | TCATAGGACAGGTCAGAGGC | ||
tcDNA +128+147 | +128+147 | TAGGACAGGTCAGAGGCAAG | |||
tcDNA +131+150 | +131+150 | tcDNA_R3 | GACAGGTCAGAGGCAAGCTT | ||
tcDNA +134+153 | +134+153 | AGGTCAGAGGCAAGCTTGAC | |||
DS | tcDNA +5−15 | +5−15 | tcDNA_DS | CCAAGgttagtgtcaagcat | |
tcDNA +3−18 | +3−18 | AAGgttagtgtcaagcatat | |||
tcDNA +1-20 | +1-20 | Ggttagtgtcaagcatatct | |||
PMO | R1 | PMO +36+60 | +36+60 | PMO_R1a | GAACAGCTGCAGAACAGGAGACAAC |
PMO +43+67 | +43+67 | PMO_R1b | GCAGAACAGGAGACAACAGTTGAAT | ||
R2 | PMO +69+98 | +69+98 | PMO_R2 | GAAATGTTAAAGGATTCAACACAAT | |
R3 | No PMO for this region | ||||
DS | PMO +9−16 | +9−16 | PMO_DS | GAAACCAAGgttagtgtcaagcata | |
MOE | MOE +9+26 | +9+26 | TTCAGATTCAGTGGGATGTG | ||
R1 | MOE +28+45 | +28+45 | MOE_R1 | GGTTCAAGAACAGCTGCA | |
R2 | MOE +79+98 | +79+98 | MOE_R2 | TCAACACAATGGCTGGAAGCT | |
MOE +83+102 | +83+102 | CAACACAATGGCTGGAAGCT | |||
MOE +84+103 | +84+103 | TCAACACAATGGCTGGAAGC | |||
R3 | MOE +135+154 | +135+154 | MOE_R3 | TCAGAGGCAAGCTTGACTCA | |
DS | MOE +5−15 | +5−15 | MOE_DS | CCAAGgttagtgtcaagcat |
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Doisy, M.; Vacca, O.; Fergus, C.; Gileadi, T.; Verhaeg, M.; Saoudi, A.; Tensorer, T.; Garcia, L.; Kelly, V.P.; Montanaro, F.; et al. Networking to Optimize Dmd exon 53 Skipping in the Brain of mdx52 Mouse Model. Biomedicines 2023, 11, 3243. https://doi.org/10.3390/biomedicines11123243
Doisy M, Vacca O, Fergus C, Gileadi T, Verhaeg M, Saoudi A, Tensorer T, Garcia L, Kelly VP, Montanaro F, et al. Networking to Optimize Dmd exon 53 Skipping in the Brain of mdx52 Mouse Model. Biomedicines. 2023; 11(12):3243. https://doi.org/10.3390/biomedicines11123243
Chicago/Turabian StyleDoisy, Mathilde, Ophélie Vacca, Claire Fergus, Talia Gileadi, Minou Verhaeg, Amel Saoudi, Thomas Tensorer, Luis Garcia, Vincent P. Kelly, Federica Montanaro, and et al. 2023. "Networking to Optimize Dmd exon 53 Skipping in the Brain of mdx52 Mouse Model" Biomedicines 11, no. 12: 3243. https://doi.org/10.3390/biomedicines11123243