TY - JOUR
T1 - Targeted SMN exon skipping
T2 - A useful control to assess in vitro and in vivo splice-switching studies
AU - Flynn, Loren L.
AU - Mitrpant, Chalermchai
AU - Adams, Abbie
AU - Pitout, Ianthe L.
AU - Stirnweiss, Anja
AU - Fletcher, Sue
AU - Wilton, Steve D.
PY - 2021/5
Y1 - 2021/5
N2 - The literature surrounding the use of antisense oligonucleotides continues to grow, with new disease and mechanistic applications constantly evolving. Furthermore, the discovery and advancement of novel chemistries continues to improve antisense delivery, stability and effectiveness. For each new application, a rational sequence design is recommended for each oligomer, as is chemistry and delivery optimization. To confirm oligomer delivery and antisense activity, a positive control AO sequence with well characterized target-specific effects is recommended. Here, we describe splice-switching antisense oligomer sequences targeting the ubiquitously expressed human and mouse SMN and Smn genes for use as control AOs for this purpose. We report two AO sequences that induce targeted skipping of SMN1/SMN2 exon 7 and two sequences targeting the Smn gene, that induce skipping of exon 5 and exon 7. These antisense sequences proved effective in inducing alternative splicing in both in vitro and in vivo models and are therefore broadly applicable as controls. Not surprisingly, we discovered a number of differences in efficiency of exon removal between the two species, further highlighting the differences in splice regulation between species.
AB - The literature surrounding the use of antisense oligonucleotides continues to grow, with new disease and mechanistic applications constantly evolving. Furthermore, the discovery and advancement of novel chemistries continues to improve antisense delivery, stability and effectiveness. For each new application, a rational sequence design is recommended for each oligomer, as is chemistry and delivery optimization. To confirm oligomer delivery and antisense activity, a positive control AO sequence with well characterized target-specific effects is recommended. Here, we describe splice-switching antisense oligomer sequences targeting the ubiquitously expressed human and mouse SMN and Smn genes for use as control AOs for this purpose. We report two AO sequences that induce targeted skipping of SMN1/SMN2 exon 7 and two sequences targeting the Smn gene, that induce skipping of exon 5 and exon 7. These antisense sequences proved effective in inducing alternative splicing in both in vitro and in vivo models and are therefore broadly applicable as controls. Not surprisingly, we discovered a number of differences in efficiency of exon removal between the two species, further highlighting the differences in splice regulation between species.
KW - Antisense oligonucleotide
KW - Cell penetrating peptide
KW - Morpholino
KW - Positive control
KW - Survival motor neuron
UR - http://www.scopus.com/inward/record.url?scp=85106557395&partnerID=8YFLogxK
U2 - 10.3390/biomedicines9050552
DO - 10.3390/biomedicines9050552
M3 - Article
C2 - 34069072
AN - SCOPUS:85106557395
SN - 2227-9059
VL - 9
JO - Biomedicines
JF - Biomedicines
IS - 5
M1 - 552
ER -