Genetically targeted chemical assembly of polymers specifically localized extracellularly to surface membranes of living neurons

Abstract

Multicellular biological systems, particularly living neural networks, exhibit highly complex organization properties that pose difficulties for building cell-specific biocompatible interfaces. We previously developed an approach to genetically program cells to assemble structures that modify electrical properties of neurons in situ, opening up the possibility of building minimally invasive cell-specific structures and interfaces. However, the efficiency and biocompatibility of this approach were challenged by limited membrane targeting of the constructed materials. Here, we design a method for highly localized expression of enzymes targeted to the plasma membrane of primary neurons, with minimal intracellular retention. Next, we show that polymers synthesized in situ by this approach form dense extracellular clusters selectively on the targeted cell membrane and that neurons remain viable after polymerization. Last, we show generalizability of this method across a range of design strategies. This platform can be readily extended to incorporate a broad diversity of materials onto specific cell membranes within tissues and may further enable next-generation biological interfaces.

Exploring Present and Future Directions in Nano-Enhanced Optoelectronic Neuromodulation

“…These insights indicate the potential of harnessing the inherent properties of neurons and glial cells to create dynamic, adaptive nanostructured interfaces, potentially eliminating the necessity for genetic modifications. This concept is further reinforced by groundbreaking work in the genetically targeted chemical assembly , in neurons, initiating a new paradigm in neural engineering aimed at enhancing the interaction between electrodes and neural tissue in vivo, thus improving therapeutic outcomes.…”

Using Chemistry To Recreate the Complexity of the Extracellular Matrix: Guidelines for Supramolecular Hydrogel–Cell Interactions

“…Synthetic chemistry has enabled the rational design of biomaterials, able to influence cells on a larger scale. Although these two fields have evolved separately, there are several calls to merge progress, − including a very recent report by DeForest and Anseth et al, toward the creation of living materials. , Here, one can envision the chemical design of a scalable hydrogel, which can be manipulated by a cell. Recent collaborative work by Kietz and Rosales lab shows this is possible. , Further innovations can leverage the synthetic material to create the environment and 3D shape, while the living cells provide the biological complexity and production of biological molecules.…”

In Situ Synthesis and Assembly of Functional Materials and Devices in Living Systems

“…Likewise, specific subcellular locations or neuron types can be targeted using genetic engineering, like using a nuclear export signal for cytoplasmic dispersion or an LCK membrane anchor for enhanced membrane expression (Figure d-e) . A recent work from the laboratories of Karl Deisseroth and Zhenan Bao has demonstrated a strategy for achieving cell-surface localization of enzymatic reaction centers. Finally, our optogenetic polymerization technique could be expanded to genetically encoded photosensitizers that operate across different spectral regions from miniSOG’s absorption peak near 450 nm .…”