TY - JOUR
T1 - SPLICER
T2 - a highly efficient base editing toolbox that enables in vivo therapeutic exon skipping
AU - Miskalis, Angelo
AU - Shirguppe, Shraddha
AU - Winter, Jackson
AU - Elias, Gianna
AU - Swami, Devyani
AU - Nambiar, Ananthan
AU - Stilger, Michelle
AU - Woods, Wendy S.
AU - Gosstola, Nicholas
AU - Gapinske, Michael
AU - Zeballos, Alejandra
AU - Moore, Hayden
AU - Maslov, Sergei
AU - Gaj, Thomas
AU - Perez-Pinera, Pablo
N1 - We thank the DNA Services staff of the Roy J. Carver Biotechnology Center at the University of Illinois, particularly Alvaro Hernandez and Chris Wright, for their support with DNA and RNA sequencing. We thank Siva Mayandi of the Roy J. Carver Biotechnology Center Cytometry and Microscopy to Omics facility at the University of Illinois for his support with FACS. This work was supported by the National Institutes of Health grants 1U01NS122102 (T.G. and P.P.P.), 1R01NS123556 (T.G. and P.P.P.), 1R01GM141296 (T.G. and P.P.P.), 1R01GM127497 (P.P.P.), 1R01GM131272 (P.P.P.), the Muscular Dystrophy Association grant MDA602798 (T.G. and P.P.P.), the American Heart Association grant 17SDG33650087 (P.P.P.), the Parkinson\u2019s Disease Foundation grant PF-IMP-1950 (T.G. and P.P.P.) and the Simons Foundation grant 887187 (T.G. and P.P.P). J.W. was supported by the Northwestern University Clinical and Translational Science Institute, grant UL1TR001422. A.M. was supported by the National Institute of Biomedical Imaging and Bioengineering of the National Institutes of Health under Award Number T32EB019944. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
PY - 2024/12
Y1 - 2024/12
N2 - Exon skipping technologies enable exclusion of targeted exons from mature mRNA transcripts, which have broad applications in medicine and biotechnology. Existing techniques including antisense oligonucleotides, targetable nucleases, and base editors, while effective for specific applications, remain hindered by transient effects, genotoxicity, and inconsistent exon skipping. To overcome these limitations, here we develop SPLICER, a toolbox of next-generation base editors containing near-PAMless Cas9 nickase variants fused to adenosine or cytosine deaminases for the simultaneous editing of splice acceptor (SA) and splice donor (SD) sequences. Synchronized SA and SD editing improves exon skipping, reduces aberrant splicing, and enables skipping of exons refractory to single splice site editing. To demonstrate the therapeutic potential of SPLICER, we target APP exon 17, which encodes amino acids that are cleaved to form Aβ plaques in Alzheimer’s disease. SPLICER reduces the formation of Aβ42 peptides in vitro and enables efficient exon skipping in a mouse model of Alzheimer’s disease. Overall, SPLICER is a widely applicable and efficient exon skipping toolbox.
AB - Exon skipping technologies enable exclusion of targeted exons from mature mRNA transcripts, which have broad applications in medicine and biotechnology. Existing techniques including antisense oligonucleotides, targetable nucleases, and base editors, while effective for specific applications, remain hindered by transient effects, genotoxicity, and inconsistent exon skipping. To overcome these limitations, here we develop SPLICER, a toolbox of next-generation base editors containing near-PAMless Cas9 nickase variants fused to adenosine or cytosine deaminases for the simultaneous editing of splice acceptor (SA) and splice donor (SD) sequences. Synchronized SA and SD editing improves exon skipping, reduces aberrant splicing, and enables skipping of exons refractory to single splice site editing. To demonstrate the therapeutic potential of SPLICER, we target APP exon 17, which encodes amino acids that are cleaved to form Aβ plaques in Alzheimer’s disease. SPLICER reduces the formation of Aβ42 peptides in vitro and enables efficient exon skipping in a mouse model of Alzheimer’s disease. Overall, SPLICER is a widely applicable and efficient exon skipping toolbox.
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U2 - 10.1038/s41467-024-54529-y
DO - 10.1038/s41467-024-54529-y
M3 - Article
C2 - 39609418
AN - SCOPUS:85210479134
SN - 2041-1723
VL - 15
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 10354
ER -