Development of A2G80 peptide-gene complex for targeted delivery to muscle cells

October 27, 2020 0 Comments

Development of A2G80 peptide-gene complex for targeted delivery to muscle cells


  • Therapeutic methods based mostly on antisense oligonucleotides and therapeutic genes are being extensively investigated for the therapy of hereditary muscle illnesses and maintain nice promise. Nevertheless, the mobile uptake of these polyanions to the muscle cells is inefficient. Subsequently, it’s mandatory to develop more practical strategies of gene delivery into the muscle tissue.


  • The A2G80 peptide (VQLRNGFPYFSY) from the laminin α2 chain has excessive affinity for α-dystroglycan (α-DG) which is expressed on the membrane of muscle cells. On this research, we designed a peptide-modified A2G80 with oligoarginine and oligohistidine (A2G80-R9-H8), and ready peptide/plasmid DNA (pDNA) complex, to develop an environment friendly gene delivery system for the muscle tissue.


  • The peptide/pDNA complex confirmed α-DG-dependent mobile uptake of the A2G80 sequence and considerably improved gene transfection effectivity mediated by the oligohistidine sequence in C2C12 myoblast cells.
  • Additional, the peptide/pDNA complex promoted environment friendly and sustained gene expression within the Duchenne muscular dystrophy mouse fashions. The A2G80-R9-H8 peptide has the potential for use as a selected provider for focusing on muscle in gene remedy in muscular dystrophy.

Use of Alcalase within the manufacturing of bioactive peptides: A evaluation


This evaluation goals to cowl the makes use of of the commercially accessible protease Alcalase within the manufacturing of biologically energetic peptides since 2010. Immobilization of Alcalase has additionally been reviewed, as immobilization of the enzyme might enhance the ultimate response design enabling the use of extra drastic situations and the reuse of the biocatalyst.

That method, this evaluation presents the manufacturing, through Alcalase hydrolysis of completely different proteins, of peptides with antioxidant, angiotensin I-converting enzyme inhibitory, metallic binding, antidiabetic, anti-inflammatory and antimicrobial actions (amongst different bioactivities) and peptides that enhance the useful, sensory and dietary properties of meals.

Alcalase has proved to be among the many best proteases for this objective, utilizing completely different protein sources, being particularly attention-grabbing the use of the protein residues from meals trade as feedstock, as this additionally solves nature air pollution issues.

Very curiously, the bioactivities of the protein hydrolysates additional improved when Alcalase is utilized in a mixed method with different proteases each in a sequential method or in a simultaneous hydrolysis (one thing that may very well be associated to the idea of combi-enzymes), as the mix of proteases with completely different selectivities and specificities allow the manufacturing of a bigger quantity of peptides and of a smaller dimension.



Artificial peptide hydrogels as 3D scaffolds for tissue engineering


The regeneration of tissues and organs poses an immense problem due to the acute complexity within the analysis work concerned. Regardless of the tissue engineering strategy being thought-about as a promising technique for greater than twenty years, a key subject impeding its progress is the dearth of very best scaffold supplies.

Nature-inspired artificial peptide hydrogels are inherently biocompatible, and its excessive resemblance to extracellular matrix makes peptide hydrogels appropriate 3D scaffold supplies. This evaluation covers the essential facets of peptide hydrogels as 3D scaffolds, together with mechanical properties, biodegradability and bioactivity, and the present approaches in creating matrices with optimized options. Many of these scaffolds include peptide sequences which might be broadly reported for tissue restore and regeneration and these peptide sequences will even be mentioned.

Moreover, 3D biofabrication methods of artificial peptide hydrogels and the current advances of peptide hydrogels in tissue engineering will even be described to replicate the present development within the area. Within the last part, we’ll current the long run outlook within the design and growth of peptide-based hydrogels for translational tissue engineering purposes.


Examine of the Interplay of a Novel Semi-Artificial Peptide with Mannequin Lipid Membranes


  • Most linear peptides straight work together with membranes, however the mechanisms of interplay are removed from being utterly understood. Right here, we current an investigation of the membrane interactions of a designed peptide containing a non-natural, artificial amino acid. We chosen a nonapeptide that’s reported to work together with phospholipid membranes, ALYLAIRKR, abbreviated as ALY.


  • We designed a modified peptide (azoALY) by substituting the tyrosine residue of ALY with an antimicrobial azobenzene-bearing amino acid. Each of the peptides have been examined for their skill to work together with mannequin membranes, assessing the penetration of phospholipid monolayers, and leakage throughout the bilayer of massive unilamellar vesicles (LUVs) and large unilamellar vesicles (GUVs).


  • The latter was carried out in a microfluidic gadget so as to research the kinetics of leakage of entrapped calcein from the vesicles on the single vesicle degree. Each sorts of vesicles have been ready from a 9:1 (mol/mol) combination of POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho(1′-rac-glycerol). Calcein leakage from the vesicles was extra pronounced at a low focus within the case of azoALY than for ALY.


  • Elevated vesicle membrane disturbance within the presence of azoALY was additionally evident from an enzymatic assay with LUVs and entrapped horseradish peroxidase. Molecular dynamics simulations of ALY and azoALY in an anionic POPC/POPG mannequin bilayer confirmed that ALY peptide solely interacts with the lipid head teams. In distinction, azoALY penetrates the hydrophobic core of the bilayers inflicting a stronger membrane perturbation as in contrast to ALY, in qualitative settlement with the experimental outcomes from the leakage assays.


Upkeep and differentiation of human ES cells on polyvinylidene fluoride scaffolds immobilized with a vitronectin-derived peptide


Polyvinylidene fluoride (PVDF) is biocompatible, straightforward to fabricate, and has piezoelectric properties; it has been used for many biomedical purposes together with stem cell engineering. Nevertheless, long-term cultivation of human embryonic stem cells (hESCs) and their differentiation towards cardiac lineages on PVDF haven’t been investigated.

Herein, PVDF nanoscaled membrane scaffolds have been fabricated by electrospinning; a vitronectin-derived peptide-mussel adhesive protein fusion (VNm) was immobilized on the scaffolds. hESCs cultured on the VNm-coated PVDF scaffold (VNm-PVDF scaffold) have been stably expanded for greater than 10 passages whereas sustaining the expression of pluripotency markers and genomic integrity.

Underneath cardiac differentiation situations, hESCs on the VNm-PVDF scaffold generated extra spontaneously beating colonies and confirmed the upregulation of cardiac-related genes, in contrast with these cultured on Matrigel and VNm alone. Thus, VNm-PVDF scaffolds could also be appropriate for the long-term tradition of hESCs and their differentiation into cardiac cells, thus increasing their utility in regenerative medication.

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