Trifluoroacetic acid tfa

DSPE-mPEG₂₀₀₀-NH₂ was provided by Shanghai ToYong Bio. Tech. Inc. (Shanghai, China). Boc-γ-Glu (OtBu)-Cys (Trt)-Gly [Boc-γ-GSH] was from Nanjing TG peptide Biotechnology Co., LTD (Nanjing, China). Paclitaxel (PTX) was bought from Jiangsu Aikang Biomedical Research and Development Co. Ltd. (Nanjing, China). Trifluoroacetic acid (TFA) was purchased from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). Dichloromethane (super dry solvent, with molecular sieves, water ≤ 30 ppm) was bought from J&K Scientific, Inc. (Beijing, China). Triisopropylsilane (97.5%) (Tis) was provided by J&K Scientific, Inc. (Beijing, China). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) was from Meryer (Shanghai) Chemical Technology Co., Ltd. (Shanghai, China). N-Hydroxy succinimide (NHS) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). DiR iodide [1-1-dioctadecyl-3,3,3,3-tetramethlindotricarboc-yanine iodide] (DiR) was purchased from AAT BioQuest (Sunnyvale, CA, USA). Acetonitrile (HPLC grade) was from Fisher Scientific (Pittsburgh, PA, USA). Paclitaxel injection (PTX injection) was obtained from Beijing Union Pharmaceutical Factory (Beijing, China). Deionized water was used during the experiments.

The rhizome of P. cyrtonema Hua. was obtained from Jinzhai County, China, and authenticated by Prof. Ruonan Xie, Anhui University of Chinese Medicine. Sigma Chemical Company provided dextrans of varying molecular weights for the experiment. A variety of sugars, including L-rhamnose (Rha), D-glucose (Glc), D-fructose (Fru), L-arabinose (Ara), D-galactose (Gal), D-xylose (Xyl), D-mannose (Man), and D-galacturonic acid (GalA) were commercially available purchased. The Millipore system provided distilled water. Kuer Co., Ltd. provided Sephadex G-200 and DEAE cellulose-52. Anthrone, 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide, Trifluoroacetic acid (TFA), 3-phenyl phenol, Sodium tetraborate, and Dimethyl sulfoxide were provided by Shanghai Macklin Biochemical Co., Ltd. N-Hexadecane-D34 (C16H34) was purchased from Aladdin industrial corporation. Anhydrous ethanol was bought from Jiangsu Qiangsheng Functional Chemistry Co., Ltd. Starter 3100 pH meter was obtained from Aarhus Instruments (Changzhou) Co., Ltd. SHIMADZU spectrophotometer was bought from Shimadzu International Trade (Shanghai) Co., Ltd. The GC–MS was obtained from Brook Dalton Company.

All plasmids used in this study were listed in Supplementary Table S1. Plasmid pmNG was prepared for TSP^scp/pDNA characterization. Plasmids of pCas12f containing dUn1Cas12f-VPR fusion and pCas12a containing dLbCas12a-VPR fusion were used for gene activation. Plasmid pCas9 (Addgene #80975), parental plasmids of PP Cas9 or PP Cas9-Cre, and minicircle DNAs of MC Cas9 or MC Cas9-Cre were used for genome editing. The PP Cas9 expressing 3xHA-Cas9 and PP Cas9-Cre expressing Cas9-Cre were constructed by standard PCR and Gibson Assembly. Plasmids expressing single guide RNA (sgRNA) were constructed by ligating the corresponding annealed oligos to the basic plasmid under the human U6 promoter (GENEWIZ, China). SgRNA sequences listed in Supplementary Table S2 were designed through https://www.benchling.com/.
Both the PP vector and the E. coli strain ZYCY10P3S2T were kindly provided by Penghui Zhou Lab.SCP (AC-TGSTQHQ-CG, disulfide bridge 2–10) was purchased from Sangon Biotech (Shanghai). Tween 85, PEI 2K, and PEI 25K were purchased from Sigma Aldrich (Shanghai). N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDCI), 4-dimethylamiopryidine (DMAP) and trifluoroacetic acid (TFA) were obtained from Macklin (Shanghai). Ditertbutyl decarbonate, 3,3’-dithiodipropionic acid (DTDP) and DTT were obtained from Aladdin (Shanghai).