Aplf clover configurator Activator#
In some embodiments, the payload comprises: an epigenetic modifier, e.g., a molecule that modifies DNA or chromatin component, e.g., a molecule that modifies a histone, e.g., an epigenetic modifier described herein, e.g., in Section VI a transcription factor, e.g., a transcription factor described herein, e.g., in Section VI a transcriptional activator domain an inhibitor of a transcription factor, e.g., an anti-transcription factor antibody, or other inhibitors a small molecule an antibody an enzyme an enzyme that interacts with DNA, e.g., a helicase, restriction enzyme, ligase, or polymerase and/or a nucleic acid, e.g., an enzymatically active nucleic acid, e.g., a ribozyme, or an mRNA, siRNA, of antisense oligonucleotide. In some embodiments, said composition comprises a payload, e.g., a payload described herein, e.g., in Section VI, e.g., in Table VI-1, VI-2, VI-3, VI-4, VI-5, VI-6, or VI-7. In other embodiments, said Cas9 molecule is an eiCas9 molecule. In some embodiments, said Cas9 molecule is an eaCas9 molecule. In some embodiments, the composition further comprises a Cas9 molecule, e.g., an eaCas9 or an eiCas9 molecule. In another aspect, the disclosure features a composition, e.g., pharmaceutical composition, comprising a gRNA molecule described herein.
In one aspect, the disclosure features a gRNA molecule comprising a targeting domain which is complementary with a target sequence from a target nucleic acid disclosed herein, e.g., a sequence from: a gene or pathway described herein, e.g., in Section VIIB, e.g., in Table VII-13, VII-14, VII-15, VII-16, VII-17, VII-18, VII-19, VII-20, VII-21, VII-22, VII-23, VII-24, VII-25, IX-1, IX-1A, IX-2, IX-3, XIV-1, or Section VIII. Depending on the Cas9 molecule/gRNA molecule complex used specific editing or the delivery of a payload can be effected. Methods and compositions disclosed herein, e.g., a Cas9 molecule complexed with a gRNA molecule, can be used to target a specific location in a target DNA. Targeted gene regulation based on the CRISPR/Cas system uses an enzymatically inactive Cas9 (also known as a catalytically dead Cas9). The CRISPR/Cas system has also been used for gene regulation including transcription repression and activation without altering the target sequence. The introduction of site-specific double strand breaks (DSBs) allows for target sequence alteration through one of two endogenous DNA repair mechanisms-either non-homologous end-joining (NHEJ) or homology-directed repair (HDR). Recently, the CRISPR/Cas system has been adapted for genome editing in eukaryotic cells. The Cas9 protein cleaves and thereby silences the viral target. That RNA, which contains sequence complimentary to the viral genome, mediates targeting of a Cas9 protein to the sequence in the viral genome. RNA is transcribed from a portion of the CRISPR locus that includes the viral sequence.
Upon exposure to a virus, short segments of viral DNA are integrated into the CRISPR locus. BACKGROUND OF TILE INVENTIONĬRISPRs (Clustered Regularly Interspaced Short Palindromic Repeats) evolved in bacteria as an adaptive immune system to defend against viral attack. The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The invention relates to CRISPR-related methods and components for editing of, or delivery of a payload to, a target nucleic acid sequence.
The contents of each of which are hereby incorporated by reference in their entirety. The present application claims the benefit of U.S.
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