Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC), a novel method for genome editing in primary human cells, has shown promising results. Previous studies have demonstrated efficient genome editing using PERC in T cells with Cas9, Cas12a, and an adenine base editor, as well as in B cells and NK cells using Cas9. These experiments utilized the INF7TAT peptide variant A5K, which was originally identified through screening for lytic activity in red blood cells, serving as a proxy for endosomal escape.

Further screening of INF7TAT variants in T cells revealed several additional activating substitutions (G1K, G20L, and Y22N), which have been incorporated into a single peptide, INF7TAT-P55 (hereafter referred to as P55). Like the A5K variant, P55 supports efficient genome editing in T cells with high yields and demonstrates suitability for CD34+ hematopoietic stem and progenitor cells (HSPCs). Both A5K and P55 have been effective in facilitating PERC in HSPCs; however, P55 is preferred in this protocol due to its slightly enhanced editing rates in these cells.

SEQUENCE: H-Lys-Leu-Phe-Glu-Ala-Ile-Glu-Gly-Phe-Ile-Glu-Asn-Gly-Trp-Glu-Gly-Met-Ile-Asp-Leu-Trp-Asn-Gly-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-OH

ONE-LETTER SEQUENCE: KLFEAIEGFIENGWEGMIDLWNGYGRKKRRQRR

MOLECULAR FORMULA: C₁₈₂H₂₈₂N₅₆O₄₈S

MOLECULAR WEIGHT: 4054.84

STORAGE CONDITIONS: -20 ± 5 °C

CAS REGISTRY NUMBER:

SYNONYMS:

RESEARCH AREA: Immunomodulation

SKU(s): IMMO-008A, IMMO-008B

KEYWORDS: delivery, CRISPR, Cas9, Cas12a, base editing, genome editing, CAR-T

REFERENCES:

1. Foss, D. V. et al. Peptide-mediated delivery of CRISPR enzymes for the efficient editing of primary human lymphocytes. Nat. Biomed. Eng. 7, 647–660 (2023).