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Additional file 1: Table S1. The sequences of DNA oligos. Figure S1. SDS-PAGE gel electrophoresis (12%) was applied to illustrate the efficiency of the loading, before and after adding AGu@PEG1500-PMO at different concentrations (including 30, 60, 120, and 240 nM) of Cas9. Cas9 concentrations were quantified with image j software. Figure S2. SDS-PAGE gel electrophoresis (12%) of a) RNP released from RNP@AGu@PEG1500-PMO at 0, 2, 4, 6, 12, and 24 h (lane 1-6), and known free RNP samples (lane a-d). Figure S3. SDS-PAGE (12%) of purified Cas9. Figure S4. Agarose gel electrophoresis (1%) of purified sgRNA. Figure S5. a) The fluorescence microscopy images with GFP filter, and b) bright field fluorescence microscopy images of the colony formed by expansion of single cell for 16, 48, and 120 h. Scale bar: 200 µm. Figure S6. Agarose gel electrophoresis (1%) of Cas9 activity assay using GFP-PCR product from pLenti6.3-To-V5-Dest- GFP (645 bp) as substrate, a) released-RNP from RNP@AGu@PEG1500-PMO complex digest the PCR product at 7.5 (lane 1), b) free-RNP, and c) digest the PCR product at 7.5 and 5 pH (lane 4 and lane 5, respectively).

Phosphonic acid functionalized KIT-6 confined ionic liquid (IL = 1-butyl-3-methylimidazolium tetrafluoroborate [BMIm][BF4]) catalyzed the one-pot condensation reaction of iminochromenes and salicylaldehydes with different primary alcohols to achieve the corresponding 4-alkoxy-5H-chromen[2,3-d]pyrimidines under solvent-free conditions and in high yields. This efficient nanocatalyst can be recovered for at least five reaction runs without significant loss of either activity or confined IL.