Zhao Z, Zhang H, Li W, Wang Y, Wang Y, Yang H, Yin L, Liu X (2025) Guanidyl-rich α-helical polypeptide enables efficient cytosolic pro-protein delivery and CRISPR-Cas9 genome editing. J Mater Chem B doi: 10.1039/d4tb02009j PMID: 39760520
Objective: To develop an efficient strategy via cationic alpha-helical polypeptide-mediated anionic proprotein delivery.
Summary: The protein was reversibly modified with adenosine triphosphateviadynamic covalent chemistry to prepare an anionic proprotein (A-protein) with abundant phosphate groups. A guanidyl-decorated a-helical polypeptide (LPP) was employed not only to encapsulate A-protein through electrostatic attraction and hydrogen bonding, forming stable nanocomplexes, but also to enhance cell membrane penetration due to its rigid alpha-helical conformation. Consequently, this strategy mediated the effective delivery of various proteins with different isoelectric points and molecular weights, including a-chymotrypsin, bovine serum albumin, ribonuclease A, cytochromeC, saporin, horseradish peroxidase, b-galactosidase, and anti-phospho-Akt, into cancer cells.
Usage: Cytosolic delivery: Saporin, was employed to evaluate LPP-mediated cytosolic delivery and its cancer cell-killing efficacy. The LPP/A-saporin nocomplexes (NCs) caused significant toxicity, with an IC50 value of 0.4ug/mL. In vivo antitumor efficiency of LPP/A-saporin NCs: LPP/A-saporin NCs were administrated by intratumoral injection in HeLa xenograft tumor-bearing mice and their antitumor efficacy was evaluated. When the tumor volume reached B50 mm3, the mice were randomly divided into three groups (6 mice per group) and intratumorally injected with PBS, free saporin, or LPP/A-saporin NCs at 0.1 mg saporin per kg on days 0 and 2.
Related Products: Saporin (Cat. #PR-01)