Abstract

Genome engineering for materials synthesis is a promising avenue for manufacturing materials with unique properties under ambient conditions. Biomineralization in diatoms, unicellular algae that use silica to construct micron-scale cell walls with nanoscale features, is an attractive candidate for functional synthesis of materials for applications including photonics, sensing, filtration, and drug delivery. Therefore, controllably modifying diatom structure through targeted genetic modifications for these applications is a very promising field. In this work, we used gene knockdown in Thalassiosira pseudonana diatoms to create modified strains with changes to structural morphology and linked genotype to phenotype using supervised machine learning. An artificial neural network (NN) was developed to distinguish wild and modified diatoms based on the SEM images of frustules exhibiting phenotypic changes caused by a specific protein (Thaps3_21880), resulting in 94% detection accuracy. Class activation maps visualized physical changes that allowed the NNs to separate diatom strains, subsequently establishing a specific gene that controls pores. A further NN was created to batch process image data, automatically recognize pores, and extract pore-related parameters. Class interrelationship of the extracted paraments was visualized using a multivariate data visualization tool, called CrossVis, and allowed to directly link changes in morphological diatom phenotype of pore size and distribution with changes in the genotype.

Highlights

  • Living organisms can construct complex three-dimensional structures from inorganic materials at ambient temperature, pressure, and pH in aqueous conditions, yet the complexity and precision of this synthesis rival modern industrial methods

  • With regard to inspected Thaps3_21880 modification, we demonstrated a neural network (NN) that can identify wild and modified diatoms with 94% accuracy

  • Two independent clonal knockdown lines were selected for nourseothricin resistance, and confirmed by polymerase chain reaction (PCR) (Supplementary materials Fig. S1 and Methods section for details).[40]

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Summary

Introduction

Living organisms can construct complex three-dimensional structures from inorganic materials at ambient temperature, pressure, and pH in aqueous conditions, yet the complexity and precision of this synthesis rival modern industrial methods. We used image processing[33,36] and machine learning classification algorithms (artificial NNs)[37,38] to screen for genes that affect diatom phenotype and to distinguish diatoms with wild type and modified morphologies.

Results
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