N n n n tetramethylethylenediamine

Hydrogenated soybean phospholipids (HSPC), cholesterol (Chol), and distearoyl phosphoethanolamine-PEG₂₀₀₀-Acrylamide (DSPE-PEG₂₀₀₀-ACA) were purchased from Shanghai Ponsure Biotech, Inc. Four-arm poly(ethylene glycol) amine (Tetra-PEG-amine) was provided by Xiamen Sinopeg company. Dibenzocyclooctyne-acid (DBCO acid, ≥95%) was supplied by Xi’an ruixi Biological Technology Co., Ltd. O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU, 99%) and glutathione (GSH, 99%) were purchased from J&K Scientific Ltd. N,N-diisopropylethylamine (≥99%, DIEA), N,N’-methylenebisAcrylamide (99%, MBAA), N,N,N’,N’-tetramethylethylenediamine (99%, TEMED), and anhydrous N,N-dimethylformamide (99.8%, DMF) were provided by Sigma. Ammonium persulfate (99.99%, APS) and 1,11-diazido-3,6,9-trioxaundecane (≥95%, N₃-PEG₃-N₃) were supplied by Aladdin. Acrylamide (99%, AM) was purchased from Adamas. Ethyl ether (≥99.7%, Et₂O) and chloroform (≥99%, CHCl₃) were provided by Sinopharm chemical reagent Co., Ltd. Ammonium sulfate (≥99%, (NH₄)₂SO₄) and PBS buffer (10×, pH 7.4) were supplied by Sangon Biotech company. The PBS buffer used in all experiments was diluted ten times. Deionized water was used to prepare all aqueous solutions. Other reagents and solvents were used as received.

All synthetic oligonucleotides were purchased from Eurofins. PCR primers were desalted, DNAzymes were purified by Eurofins proprietary High Purity Salt Free reverse phase cartridge purification system and short RNA substrates were purified by HPLC. RNase-free ultrapure water was produced using a Milli-Q Advantage A10 Water Purification System equipped with a BioPak® Polisher from Merck Millipore. Phusion High-Fidelity PCR kit and TranscriptAid T7 High Yield Transcription kit were from Life Technologies. 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), sodium chloride, manganese chloride tetrahydrate, formamide, bromophenol blue, 40% 19:1 acrylamide/bis-acrylamide, N,N,N′,N′-tetramethylethylenediamine, ammonium persulfate, urea, agarose, ethidium bromide and phenol:chloroform:isoamyl alcohol 25:24:1 pH 8.0 was purchased from Sigma-Aldrich. Ethylenediaminetetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) were from Scharlau. Boric acid was from VWR. Klorrent bleach was from Nilfisk. The nucleic acid dyes SYTO 61, PO-PRO 1, DRAQ5, Hoechst 33258, DAPI, Propidium iodide and PicoGreen were all from Life Technologies. GelRed was from Biotium. ethidium bromide was from Sigma.

The tissue-mimetic hydrogels were prepared as follows: Acrylamide, N, N methylene-bis-Acrylamide, ammonium persulphate (APS) and N, N, N′, N′-tetramethyl -ethylenediamine (TEMED) were purchased from Sigma-Aldrich. A solution containing Acrylamide monomers, crosslinker N, N methylene-bis-Acrylamide, ammonium persulphate and N, N, N′, N′- tetramethylethylenediamine (TEMED) was prepared. Then various concentrations of GNRs dispersed in PBS were added to the prepared solution and mixed thoroughly. The prepared solution was poured inside a micromachined glass mold to make the desired thickness (300 μm) of the PolyAcrylamide (PAAm) hydrogels. Hydrogels with different stiffness were prepared to investigate the effect of the stiffness on the heat generation. The ratio of Acrylamide%: bis-Acrylamide% was varied (6:0.06, 10:0.1 and 10:0.3) in the process of gel making to adjust the hydrogel stiffness and porosity as reported elsewhere^{70 (link),71 (link)}.