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The invention also provides for a conjugate comprising the oligomer according to the invention as herein described, and at least one non-nucleotide or non-polynucleotide moiety covalently attached to said oligomer. In one embodiment, random therapeutic molecules comprising nucleotide sequences (e.g., oligomers) targeting certain regions of pre-mRNA or mRNA encoding MAPT, BASP1, or APP are prepared to test their toxicities. Another example of a target nucleic acid sequence of the oligomers is BASP1 pre-mRNA or BASP1 mRNA. Sequence score is a mathematical calculation determined for all oligomers and is based on the percent of G and C nucleotides, or analogs thereof, within a given oligomer sequence. The sequence score of the selected oligomers were calculated for further studies. A further example of a target nucleic acid sequence of the oligomers is APP pre-mRNA or APP mRNA. The nucleotides of the oligomer of the invention or contiguous nucleotides sequence thereof can be coupled together via linkage groups.
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Brain tissue (right, frontal cortical region) was collected on dry ice or RNAlater for site porno drug concentration analysis and Tau qPCR respectively at multiple time points following dosing, e.g., 1 week through 16 weeks post-dosing. Tau gene was deleted, were used to assess pharmacodynamic endpoints and tissue drug concentrations. The BASP1 gene encodes a membrane bound protein with several transient phosphorylation sites and PEST motifs. In a pathologic state associated with disease, MAPT is also known as neurofibrillary tangle protein or paired helical filament-tau (PHF-tau). Natural variants of the MAPT gene product are known. In humans, the gene for APP is located on chromosome 21 and contains 18 exons spaning 290 kilobases. Amyloid precursor protein (APP) is an integral membrane protein expressed in many tissues and concentrated in the synapses of neurons. BASP1 is also known as 22 kDa neuronal tissue-enriched acidic protein, neuronal axonal membrane protein NAP-22, NAP22, CAP-23, NAP-22, CAP23, or Neuronal Tissue-Enriched Acidic Protein.
In certain embodiments, the oligomers target a pre-mRNA or mRNA encoding brain acid soluble protein 1 (BASP1). In certain embodiments, the oligomers target microtubule-associated protein tau (MAPT). A number of molecules (e.g., oligomers) are designed to target the 3' UTR of MAPT pre-mRNA. The therapeutic molecules can be prepared with carriers that protect against degradation or immediate elimination from the body, including implants or microcapsules with controlled release properties. As discussed in Example 2, the effect of small molecules on primary neuronal spontaneous calcium oscillations will be measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Effect of oligomers on primary neuronal spontaneous calcium oscillations was measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Therapeutic proteins that reduce calcium oscillations to levels below 70% of control will be expected to negatively impact CNS network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro.
