TY - JOUR
T1 - Spell Checking Nature
T2 - Versatility of CRISPR/Cas9 for Developing Treatments for Inherited Disorders
AU - Wojtal, Daria
AU - Kemaladewi, Dwi U.
AU - Malam, Zeenat
AU - Abdullah, Sarah
AU - Wong, Tatianna W.Y.
AU - Hyatt, Elzbieta
AU - Baghestani, Zahra
AU - Pereira, Sergio
AU - Stavropoulos, James
AU - Mouly, Vincent
AU - Mamchaoui, Kamel
AU - Muntoni, Francesco
AU - Voit, Thomas
AU - Gonorazky, Hernan D.
AU - Dowling, James J.
AU - Wilson, Michael D.
AU - Mendoza-Londono, Roberto
AU - Ivakine, Evgueni A.
AU - Cohn, Ronald D.
N1 - Funding Information:
We would like to thank Drs. Jennifer Doudna, Steve Lin, Aravinda Chakravarti, Hal Dietz, and Janet Rossant for critical insights into our studies, as well as Minggao Liang and Wilson Sung for their bioinformatics input. We also thank Drs. Feng Zhang and Rudolf Jaenisch and their laboratories for the CRISPR/Cas9 backbone constructs used in this study. We thank the R.D.C. lab members and The Centre for Applied Genomics staff for excellent technical assistance, as well as the platform for immortalization of human cells of the Institute of Myology in Paris. This work is supported by SickKids Restracomp and Eric Hani Fellowship (to D.W.), AFM-Telethon (to D.U.K), Cure CMD (to D.U.K. and R.D.C.), a tier II Natural Sciences and Engineering Research Council Canada Research Chair (436194-2013 to M.D.W), the Duchenne Children’s Trust (to R.D.C.), the Women's Auxiliary of the The Hospital for Sick Children (Chair in Clinical and Metabolic Genetics to R.D.C.), and the SickKids Foundation (to R.D.C.).
Funding Information:
We would like to thank Drs. Jennifer Doudna, Steve Lin, Aravinda Chakravarti, Hal Dietz, and Janet Rossant for critical insights into our studies, as well as Minggao Liang and Wilson Sung for their bioinformatics input. We also thank Drs. Feng Zhang and Rudolf Jaenisch and their laboratories for the CRISPR/Cas9 backbone constructs used in this study. We thank the R.D.C. lab members and The Centre for Applied Genomics staff for excellent technical assistance, as well as the platform for immortalization of human cells of the Institute of Myology in Paris. This work is supported by SickKids Restracomp and Eric Hani Fellowship (to D.W.), AFM-Telethon (to D.U.K), Cure CMD (to D.U.K. and R.D.C.), a tier II Natural Sciences and Engineering Research Council Canada Research Chair (436194-2013 to M.D.W), the Duchenne Children''s Trust (to R.D.C.), the Women''s Auxiliary of the The Hospital for Sick Children (Chair in Clinical and Metabolic Genetics to R.D.C.), and the SickKids Foundation (to R.D.C.).
Publisher Copyright:
© 2016 The American Society of Human Genetics.
PY - 2016/1/7
Y1 - 2016/1/7
N2 - Clustered regularly interspaced short palindromic repeat (CRISPR) has arisen as a frontrunner for efficient genome engineering. However, the potentially broad therapeutic implications are largely unexplored. Here, to investigate the therapeutic potential of CRISPR/Cas9 in a diverse set of genetic disorders, we establish a pipeline that uses readily obtainable cells from affected individuals. We show that an adapted version of CRISPR/Cas9 increases the amount of utrophin, a known disease modifier in Duchenne muscular dystrophy (DMD). Furthermore, we demonstrate preferential elimination of the dominant-negative FGFR3 c.1138G>A allele in fibroblasts of an individual affected by achondroplasia. Using a previously undescribed approach involving single guide RNA, we successfully removed large genome rearrangement in primary cells of an individual with an X chromosome duplication including MECP2. Moreover, removal of a duplication of DMD exons 18-30 in myotubes of an individual affected by DMD produced full-length dystrophin. Our findings establish the far-reaching therapeutic utility of CRISPR/Cas9, which can be tailored to target numerous inherited disorders.
AB - Clustered regularly interspaced short palindromic repeat (CRISPR) has arisen as a frontrunner for efficient genome engineering. However, the potentially broad therapeutic implications are largely unexplored. Here, to investigate the therapeutic potential of CRISPR/Cas9 in a diverse set of genetic disorders, we establish a pipeline that uses readily obtainable cells from affected individuals. We show that an adapted version of CRISPR/Cas9 increases the amount of utrophin, a known disease modifier in Duchenne muscular dystrophy (DMD). Furthermore, we demonstrate preferential elimination of the dominant-negative FGFR3 c.1138G>A allele in fibroblasts of an individual affected by achondroplasia. Using a previously undescribed approach involving single guide RNA, we successfully removed large genome rearrangement in primary cells of an individual with an X chromosome duplication including MECP2. Moreover, removal of a duplication of DMD exons 18-30 in myotubes of an individual affected by DMD produced full-length dystrophin. Our findings establish the far-reaching therapeutic utility of CRISPR/Cas9, which can be tailored to target numerous inherited disorders.
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UR - http://www.scopus.com/inward/citedby.url?scp=84954388645&partnerID=8YFLogxK
U2 - 10.1016/j.ajhg.2015.11.012
DO - 10.1016/j.ajhg.2015.11.012
M3 - Article
C2 - 26686765
AN - SCOPUS:84954388645
SN - 0002-9297
VL - 98
SP - 90
EP - 101
JO - American Journal of Human Genetics
JF - American Journal of Human Genetics
IS - 1
ER -