Thiol modifier c6 s s

The crRNAs were designed using the Cas13design tool^{66 (link)}. The crRNA plasmids were cloned using standard restriction-ligation cloning. Forward and reverse oligos for each spacer (IDT) were annealed and inserted into a pHR backbone using T4 DNA Ligase (NEB). The backbone was modified from Addgene (#121514). Spacer sequences and predicted efficiency score for all crRNAs can be found in Supplementary Data 1. Some crRNAs were synthesized by Synthego using solid-phase phosphoramidite chemistry to contain 3’ Um modifications with additional 5’ Cy5 labeling^{67 (link)}. A protected 3’-thiol moiety was installed using Thiol-Modifier C6 S-S (Cat# 10-1936-90E, Glen Research). Following cleavage, deprotection and isolation, the free thiol was liberated using dithiothreitol and then reacted with Cyanine5 Maleimide (Cat# 43080, Lumiprobe) following the manufacturer’s guidelines. Purity and identity were confirmed via HPLC/ESI-MS. These are listed in Supplementary Data 1. They were used in the treatment tests as described below.

Phosphoramidite reagents used for the synthesis of FNA were purchased from Glen Research (VA, United States): spacer 18 phosphoramidite (10–1918), DBCO-dT-CE phosphoramidite (10–1539), Thiol-Modifier C6 S-S (10–1936), biotin CPG (20–2993). N-terminus azide modified Aβ42 [K(N3)-Aβ42] peptide was purchased from GenScript Biotech (NJ, United States). N-(g-Maleimidobutyryloxysuccinimide ester) GMBS was purchased from Pierce Biotechnology (Grand Island, NY). Streptavidin was obtained from Sigma-Aldrich (MO, United States) and Tris-(2-Carboxyethyl) phosphine-HCl (TCEP-HCl) was purchased from Hampton Research Inc. (CA, United States). 1-(3-aminopropyl) silatrane (APS) was synthesized as previously described (Shlyakhtenko et al., 2013 (link)). β-mercaptoethanol, NHS-PEG₄-DBCO and all other reagents or solvents were purchased from Sigma–Aldrich (MO, United States).