Cell-penetrating peptides (CPPs): From delivery of nucleic acids and antigens to transduction of engineered nucleases for application in transgenesis

Abstract

Cell-penetrating peptides (CPPs) have been studied for their capacity to translocate across the lipid membrane of several cell types. In membrane translocation, these peptides can remarkably transport biologically active hydrophilic molecules, such as pharmaceuticals, nucleic acids (DNA and RNA) and even high-molecular-weight proteins, Fig. 3 into the cell cytoplasm and organelles. The development of CPPs as transduction agents includes the modification of gene and protein expression, the reprogramming and differentiation of induced pluripotent stem cells and the preparation of cellular vaccines. A relatively recent field of CPP application is the transduction of plasmid DNA vectors and CPP-fusion proteins to modify genomes and introduce new traits in cells and organisms. CPP-mediated transduction of components for genome editing is an advantageous alternative to viral DNA vectors. Engineered site-specific nucleases, such as Cre recombinase, ZFN, TALENs and CRISPR associated protein (Cas), have been coupled to CPPs, and the fused proteins have been used to permeate targeted cells and tissues. The functionally active fusion CPP-nucleases subsequently home to the nucleus, incise genomic DNA at specific sites and induce repair and recombination. This review has the objective of discussing CPPs and elucidating the prospective use of CPP-mediated transduction technology, particularly in genome modification and transgenesis.

The TAL Effector AvrBs3 from Xanthomonas campestris pv. vesicatoria Contains Multiple Export Signals and Can Enter Plant Cells in the Absence of the Type III Secretion Translocon

“…Autonomously translocating effectors present a novel class of cell-penetrating peptides (CPPs), which are often used as vehicles for the transport of cargo molecules into target cells (Rüter and Schmidt, 2017 ). CPPs include basic/amphiphilic or hydrophobic peptides which consist of usually less than 30 amino acids and autonomously translocate across biological membranes, even when present as fusion partner of a large cargo protein (Takeuchi and Futaki, 2016 ; Radis-Baptista et al, 2017 ). CPPs are either directly transported across the membrane or are delivered by endocytosis (Scharnert et al, 2013 ; Radis-Baptista et al, 2017 ).…”

“…CPPs include basic/amphiphilic or hydrophobic peptides which consist of usually less than 30 amino acids and autonomously translocate across biological membranes, even when present as fusion partner of a large cargo protein (Takeuchi and Futaki, 2016 ; Radis-Baptista et al, 2017 ). CPPs are either directly transported across the membrane or are delivered by endocytosis (Scharnert et al, 2013 ; Radis-Baptista et al, 2017 ). Both transport pathways were observed for YopM from Yersinia spp.…”

High‐efficiency protein delivery into transfection‐recalcitrant cell types

“…Viral transduction methods are more efficient, but carry the risk of insertional mutagenesis. The cellpermeant peptide mediated Cre protein transduction method offers a safe alternative (Rádisbaptista, Campelo, Jrl, Melo, & Vjf, 2017), but the high cost of expression and purification of the fusion proteins as well as protein size limitation hinder its usages.…”

Recombinant histones as an instrument for delivery of nucleic acids into eukaryotic cells

“…При образовании комплексов с нуклеиновыми кислотами такие белки/пептиды обеспечивают прямую трансдукцию в ядра клеток, благодаря наличию в своем составе белоктрансдуцирующих доменов (БТД), которые состоят из 10-30 основных аминокислотных остатков [10][11][12][13]. Примерами таких белков могут служить гистоны, естественными функциями которых являются связывание и образование прочного комплекса с молекулами ДНК в клетке.…”