Lifeboat Foundation

Circa 2017

Schizophrenia is a genetically related mental illness, in which the majority of genetic alterations occur in the non-coding regions of the human genome. In the past decade, a growing number of regulatory non-coding RNAs (ncRNAs) including microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have been identified to be strongly associated with schizophrenia. However, the studies of these ncRNAs in the pathophysiology of schizophrenia and the reverting of their genetic defects in restoration of the normal phenotype have been hampered by insufficient technology to manipulate these ncRNA genes effectively as well as a lack of appropriate animal models. Most recently, a revolutionary gene editing technology known as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9; CRISPR/Cas9) has been developed that enable researchers to overcome these challenges. In this review article, we mainly focus on the schizophrenia-related ncRNAs and the use of CRISPR/Cas9-mediated editing on the non-coding regions of the genomic DNA in proving causal relationship between the genetic defects and the pathophysiology of schizophrenia. We subsequently discuss the potential of translating this advanced technology into a clinical therapy for schizophrenia, although the CRISPR/Cas9 technology is currently still in its infancy and immature to put into use in the treatment of diseases. Furthermore, we suggest strategies to accelerate the pace from the bench to the bedside. This review describes the application of the powerful and feasible CRISPR/Cas9 technology to manipulate schizophrenia-associated ncRNA genes. This technology could help researchers tackle this complex health problem and perhaps other genetically related mental disorders due to the overlapping genetic alterations of schizophrenia with other mental illnesses.

Keywords: CRISPR/Cas9; gene editing; lncRNAs; miRNAs; non-coding RNAs; schizophrenia.

CRISPR-Cas9 is considered a revolutionary gene editing tool, but its applications are limited by a lack of methods by which it can be safely and efficiently delivered into cells. Recently, a research team from Kumamoto University, Japan, have constructed a highly flexible CRISPR-Cas9 carrier using aminated polyrotaxane (PRX) that can not only bind with the unusual structure of Cas9 and carry it into cells, but can also protect it from intracellular degradation by endosomes.

Clustered regularly interspaced short palindromic repeats (CRISPR) and their accompanying protein, CRISPR-associated protein 9 (Cas9), made international headlines a few years ago as a game-changing genome editing system. Consisting of Cas9 and strand of genetic material known as a single-guide RNA (sgRNA), the system can target specific regions of DNA and function as “molecular scissors” to make precise edits. The direct delivery of Cas9–sgRNA complexes, i.e. Cas9 ribonucleoproteins (RNPs), into the nucleus of the cell is considered the safest and most efficient way to achieve genome editing. However, the Cas9 RNP has poor cellular permeability, and thus requires a carrier molecule to transport it past the first hurdle of the cell membrane before it can get to the cell nucleus. These carriers need to bind with Cas9 RNP, carry it into the cell, prevent its degradation by intracellular organelles called “endosomes,” and finally release it without causing any changes to its structure.

In a recent paper published in the June 2022, Volume 27 of Applied Materials Today, a research team from Kumamoto University has developed a transformable polyrotaxane (PRX) carrier that can facilitate genome editing using Cas9RNP with high efficiency and usability. “While there have been some PRX-based drug carriers for nucleic acids and proteins reported before, this is the first report on PRX-based Cas9 RNP carrier. Moreover, our findings describe how to precisely control intracellular dynamics across multiple steps. This will prove invaluable for future research in this direction,” says Professor Keiichi Motoyama, a corresponding author of the paper.

EPFL scientists have developed a digital model of the fruit fly, Drosophila melanogaster, that realistically simulates the movements of the animal. The twin is a big step towards reverse engineering the neuromechanical control of animal behavior, and developing bioinspired robots.

“We used two kinds of data to build NeuroMechFly,” says Professor Pavan Ramdya at EPFL’s School of Life Sciences. “First, we took a real fly and performed a CT scan to build a morphologically realistic biomechanical model. The second source of data were the real limb movements of the fly, obtained using pose estimation software that we’ve developed in the last couple of years that allow us to precisely track the movements of the animal.”

Ramdya’s group, working with the group of Professor Auke Ijspeert at EPFL’s Biorobotics Laboratory, has published a paper in Nature Methods showcasing the first ever accurate “digital twin” of the fly Drosophila melanogaster, dubbed “NeuroMechFly”.

Just as countries import a vast array of consumer goods across national borders, so living cells are engaged in a lively import-export business. Their ports of entry are sophisticated transport channels embedded in a cell’s protective membrane. Regulating what kinds of cargo can pass through the borderlands formed by the cell’s two-layer membrane is essential for proper functioning and survival.

Award-winning author and futurist Amy Webb examines the world of synthetic biology in her book “The Genesis Machine.” She sits down with Hari Sreenivasan to discuss the potential and the concerns of redesigning our lives.

Originally aired on April 28, 2022.

Continue reading “What is Synthetic Biology? How Scientists Are Rewriting Life | Amanpour and Company” »

I have created an educational guide to the adenovirus capsid! The adenovirus is one of the most frequently used types of viruses for gene therapy (along with AAV and lentivirus). It is a powerful vehicle for delivering DNA to cells in the body. But to work with adenovirus as a technology, it is important to understand its fundamental biological structure and function. This guide will help you to gain a more holistic comprehension of a particularly important part of adenovirus biology: the capsid. I made the images using PyMol.

PDF version: Guide to the Structure and Function of the Adenovirus Capsid

For this guide, I will explain the fundamental biology of adenovirus capsid proteins with an emphasis on the context of gene therapy. While the guide is meant primarily for readers with an interest in applying adenovirus to gene therapy, it will not include much discussion of the techniques and technologies involved in engineering adenoviruses for such purposes. If you are interested in learning more about adenovirus engineering, you may enjoy my review paper “Synthetic Biology Approaches for Engineering Next-Generation Adenoviral Gene Therapies” [1]. Here, I will focus mostly on the capsid of human adenovirus serotype 5 (Ad5) since it is the most commonly used type of adenovirus employed in gene therapy research, but I will occasionally describe other types of adenoviruses when necessary. Many of the presented concepts remain the same or similar across other types of adenoviruses.

Continue reading “Guide to the Structure and Function of the Adenovirus Capsid” »

Our podcast on the science and technology making the news. This week, we speak to Jennifer Doudna, the Nobel laureate who pioneered the revolutionary tool | Podcasts.

Eradicating Cancer With A Universal Preventative Cancer Vaccine — Dr. Stephen Johnston, Ph.D., ASU Biodesign Institute / Calviri

Dr. Stephen Johnston, Ph.D. (https://biodesign.asu.edu/stephen-johnston) is the Director for the Center for Innovations in Medicine (https://biodesign.asu.edu/Research/Centers/innovations-medicine), a Professor in the School of Life Sciences, and Director of the Biological Design Graduate Program at The Biodesign Institute at Arizona State University.

Continue reading “Dr. Stephen Johnston, PhD — Calviri — Cancer Eradication Via A Universal Preventative Cancer Vaccine” »

Cancer cells thrive by competing with normal cells for survival. Now, researchers are employing living bacteria to fight back against the cancer. This so-called bacteriotherapy – the deployment of bacteria to fight cancer – has sparked interest in the fields of immunotherapy and bioengineering.