Materials and methods for treatment of myotonic dystrophy type 1 (DM1) and other related disorders
The present application provides materials and methods for treating a patient with one or more conditions associated with DMPK whether ex vivo or in vivo. In addition, the present application provides materials and methods for editing and/or modulating the expression of DMPK gene in a cell by genome editing.
1. A single-molecule guide RNA comprising a spacer sequence that is an RNA sequence corresponding to SEQ ID NO: 5800, and further comprising a tracrRNA sequence.
2. The single-molecule guide RNA of claim 1 , wherein the single-molecule guide RNA:
further comprises a spacer extension region;
further comprises a tracrRNA extension region;
is chemically modified; and/or
is pre-complexed with a DNA endonuclease.
3. The single-molecule guide RNA of claim 2 , wherein the DNA endonuclease is a Cas9 endonuclease.
4. The single-molecule guide RNA of claim 3 , wherein the Cas9 endonuclease is selected from: S. pyogenes Cas9, S. aureus Cas9, N. meningitides Cas9, S. thermophilus CRISPR1 Cas9, S. thermophilus CRISPR 3 Cas9, T denticola Cas9, and variants having at least 90% homology to said endonucleases.
5. The single-molecule guide RNA of claim 4 , wherein the Cas9 endonuclease comprises one or more nuclear localization signals (NLSs), wherein at least one NLS is at or within 50 amino acids of the amino-terminus of the Cas9 endonuclease and/or at least one NLS is at or within 50 amino acids of the carboxy-terminus of the Cas9 endonuclease.
6. A DNA encoding the single-molecule guide RNA of claim 1 .