Sulfosuccinimidyl 4 n maleimidomethyl cyclohexane 1 carboxylate sulfo smcc

Res-β-glc, β-glc, bovine serum albumin (BSA), N-hydroxysuccinimide (NHS), tris(2-carboxyethyl)phosphine hydrochloride (TCEP), 2-(N-morpholino)ethanesulfonic acid (MES) buffer, and MNPs were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) was obtained from Thermo Fisher Scientific (Waltham, MA, USA). N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide (EDC) was purchased from Tokyo Chemical Industry (Tokyo, Japan). 1X phosphate-buffered saline (PBS, pH 7.4) solution (Gibco, Thermo Fisher Scientific) was used to dissolve β-glc. Distilled water from a Milli-Q water purification system was used to prepare all chemical solutions. FMDV (pan, O, and A type) antibodies were provided from the Korea Research Institute of Bioscience and Biotechnology. Inactivated antigens of serotype O (O1/Manisa) and A (A22/Iraq) were purchased from The Pirbright Institute (Surrey, UK).

Sicastar COOH (300 nm, 50 mg/mL), Sicastar-greenF
COOH (100 nm, 200 nm, 25 mg/mL), Sicastar-redF COOH (200 nm, 25 mg/mL),
Sicastar-greenF NH₂ (200 nm, 25 mg/mL), and Sicastar-redF
NH₂ (200 nm, 25 mg/mL) nanoparticles were obtained from
MicroMod. GreenF particles were used in DNA-PAINT experiments, redF
particles were used in the ADCP assay. Cetuximab (Erbitux), kanamycin,
BugBuster, benzonase were obtained from Merck. N-(3-(Dimethylamino)propyl)-N′-ethylcarbodiimide hydrochloride (EDC), MES, PBS
tablets, chloramphenicol, arabinose, were obtained from Sigma-Aldrich.
PD10 columns and HiTrap Q HP columns were purchased at GE Healthcare.
QuickChange Lightning multisite-directed mutagenesis kit was obtained
from Agilent. E. coli BL21(DE3)
was purchased at Novagen. β-d-1-Thiogalactopyranoside
was obtained from IPTG. Unnatural amino acid p-BpA was obtained from
Bachem. Desthiobiotin originates from IBA Life Sciences Zeba spin
columns (0.5 mL, 7 kDa MWCO) and sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC) were purchased
from ThermoFisher Scientific. NH₂–ODN dockings and
I1 were obtained from Integrated DNA Technologies, and IPS3 was obtained
from Eurofins. ODNs were dissolved in storage TE buffer and used fresh.
The sequences of ODNs used are listed in Table 1. Specific buffer was used for imaging. Buffer
B+: 10 mM MgCl₂, 54 mM Tris-HCl, 1 mM EDTA, 0.05 v/v% Tween
20, pH 8.

Detection of protein concentrations of VLPs-NC and VLPs-MEG3 was performed using BCA assays (Beyotime Biotechnology, Beijing, China), according to the manufacturer's instructions. A cysteine residue was added to the N-terminus of the GE11 polypeptides for crosslinking to the surface of MS2 VLPs, which were synthesized by Chinese Peptide Company (Hangzhou, China). The crosslinker reagent sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC) was obtained from Thermo Fisher Scientific (Rockford, IL). The VLPs-NC and VLPs-MEG3 were crosslinked with the GE11-Cys polypeptide according to the manufacturer's instructions. SDS-PAGE on 12% gels was performed to verify crosslinking.

Olaparib precursor (4‐(4‐fluoro‐3‐(piperazine‐1‐carbonyl)benzyl)phthalazin‐1(2H)‐one) was purchased from Ambeed. DBCO‐sulfo‐NHS (dibenzocyclooctyne‐sulfo‐N‐hydroxysuccinimidyl) ester was obtained from Click Chemistry Tools. Mal‐PEG₄‐NHS (O‐[N‐(3‐maleimidopropionyl)aminoethyl]‐O′‐[3‐(N‐succinimidyloxy)−3‐oxopropyl]triethylene glycol) ester and low molecular weight branched polyethylenimine (Mw 2,000 Da) were purchased from Merck/MilliporeSigma. AzF (4‐azido‐l‐phenylalanine) was purchased from Chem‐Impex International. Sulfo‐succinimidyl‐4‐(N‐maleimidomethyl) cyclohexane‐1‐carboxylate (sulfo‐SMCC) and lipofectamine CRISPRMAX transfection reagent (CMAX) was purchased from Thermo Fisher Scientific. Primary antibodies mouse anti‐Rad52 (F‐7, sc‐365341), rabbit anti‐lamin A/C, and mouse anti‐transferrin; and horseradish peroxidase (HRP)‐conjugated secondary antibody were obtained from Santa Cruz Biotechnology. BODIPY‐FL‐PEG₄‐DBCO was obtained from Jena Bioscience. TAT peptide (GRKKRRQRRRPQ) was obtained from GenScript. Primers used for DNA cloning were synthesized by Neoprobe (Table S1, Supporting Information). Primers used for target DNA preparation, sgRNA synthesis, and targeted deep sequencing were provided by Macrogen (Table S2, Supporting Information).