Exploiting the CRISPR/Cas9 system to study alternative splicing in vivo: application to titin.
The giant protein titin is the third most abundant protein in striated muscle. Mutations in its gene are responsible for diseases affecting the cardiac and/or the skeletal muscle. Titin has been reported to be expressed in multiple isoforms with considerable variability in the I-band, ensuring the modulation of the passive mechanical properties of the sarcomere. In the M-line, only the penultimate Mex5 exon coding for the specific is7 domain has been reported to be subjected to alternative splicing. Using the CRISPR-Cas9 editing technology, we generated a mouse model where we stably prevent the expression of alternative spliced variant(s) carrying the corresponding domain. Interestingly, the suppression of the domain induces a phenotype mostly in tissues usually expressing the isoform that has been suppressed, indicating that it fulfills (a) specific function(s) in these tissues allowing a perfect adaptation of the M-line to physiological demands of different muscles.
INSERM, U951, INTEGRARE research unit Evry, France|Généthon, Evry, France
Karine
K
Charton
INSERM, U951, INTEGRARE research unit Evry, France|Généthon, Evry, France
Laurence
L
Suel
INSERM, U951, INTEGRARE research unit Evry, France|Généthon, Evry, France|University of Evry-Val-D’Essone, Evry, France
Sara F
SF
Henriques
SEAT - SErvice des Animaux Transgéniques CNRS -TAAM -phenomin UPS44 Bâtiment G 7, rue Guy Môquet 94800 Villejuif, France.
Jean-Paul
JP
Moussu
Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
Matteo
M
Bovolenta
SEAT - SErvice des Animaux Transgéniques CNRS -TAAM -phenomin UPS44 Bâtiment G 7, rue Guy Môquet 94800 Villejuif, France.
Miguel
M
Taillepierre
SEAT - SErvice des Animaux Transgéniques CNRS -TAAM -phenomin UPS44 Bâtiment G 7, rue Guy Môquet 94800 Villejuif, France.
Céline
C
Becker
SEAT - SErvice des Animaux Transgéniques CNRS -TAAM -phenomin UPS44 Bâtiment G 7, rue Guy Môquet 94800 Villejuif, France.
Karelia
K
Lipson
INSERM, U951, INTEGRARE research unit Evry, France|Généthon, Evry, France
Isabelle
I
Richard
D017398:Alternative Splicing; D000818:Animals; D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D008297:Male; D051379:Mice; D023421:Models, Animal; D020033:Protein Isoforms / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D012518:Sarcomeres / Q000378:metabolism
D020033:Protein Isoforms; D011494:Protein Kinases; C579730:titin protein, mouse