Genetic Engineering Publications - GEG Tech top picks

A recent study published in the journal Nature Medicine, led by researchers James Riley, PhD, a professor of Microbiology at the Perelman School of Medicine at the University of Pennsylvania, and Todd Allen, PhD, a professor of Medicine at Harvard Medical School and Group Leader at the Ragon Institute of MGH, MIT and Harvard, describes a new Dual CAR T cell immunotherapy that can help fight HIV infection.

An innovative combination therapy can force malignant breast cancer cells to turn into fat cells. This can be used to prevent the formation of metastases in mice, as researchers at the University of Basel’s Department of Biomedicine recently reported in the journal Cancer Cell.

Here, the authors review historical cases of CCR5-edited cell-based therapies, current clinical trials and future benefits and challenges associated with this technology.

Using CRISPR/Cas9 technology, researchers have devised a method to deliver a CAR gene to a specific locus, TRAC, in T cells. This targeted approach yielded therapeutic cells that were more potent even at low doses; in a mouse model of acute lymphoblastic leukemia, they outperformed CAR T cells created with a randomly integrating retroviral vector.

This review seeks to summarize and discuss the application of clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems (Cas) for genome editing, also called genome surgery, in the field of ophthalmology.

Progressive multifocal leukoencephalopathy (PML) is a debilitating disease resulting from infection of oligodendrocytes by the JC polyomavirus (JCPyV) and with no anti-viral therapeutic available against JCPyV infection.

In this study, the authors show that the CRISPR/Cas9 system can restrict the JCPyV life cycle in cultured cells with the use of CRISPR/Cas9 to target the noncoding control region and the late gene open reading frame of the JCPyV genome. They found significant inhibition of virus replication and viral protein expression in cells recipient of Cas9 together with JCPyV-specific single-guide RNA delivered prior to or after JCPyV infection.

The repair of a point mutation can be facilitated by combined activity of a single-stranded oligonucleotide and a CRISPR/Cas9 system. In this study, the scientists provide a detailed analysis of allelic variance at and surrounding the target site. In one particular case, they report sequence alteration directed by a distinct member of the same gene family. Their data suggests that single-stranded DNA molecules may influence DNA junction heterogeneity created by CRISPR/Cas9.

In this study, the scientists  have assessed a novel panel of Morpholino ASOs with the goal of optimizing E1 ASO activity. Screening for efficacy in the SMNΔ7 mouse model, a single ASO variant was more active in vivo compared with the original E1MO-ASO. Sequence variant eleven (E1MOv11) consistently showed greater efficacy by increasing the lifespan of severe Spinal Muscular Atrophy mice after a single intracerebroventricular injection in the central nervous system, exhibited a strong dose-response across an order of magnitude, and demonstrated excellent target engagement by partially reversing the pathogenic SMN2 splicing event. They conclude that Morpholino modified ASOs are effective in modifying SMN2 splicing and have the potential for future Spinal Muscular Atrophy clinical applications.

This article reports the application of RNA nanotechnology for specific and efficient delivery of anti-miRNA seed-targeting sequence to block the growth of prostate cancer in mouse models. Utilizing the thermodynamically ultra-stable three-way junction of the pRNA of phi29 DNA packaging motor, RNA nanoparticles were constructed by bottom-up self-assembly containing the anti-prostate-specific membrane antigen (PSMA) RNA aptamer as a targeting ligand and anti-miR17 or anti-miR21 as therapeutic modules. The 16 nm RNase-resistant and thermodynamically stable RNA nanoparticles remained intact after systemic injection in mice and strongly bound to tumors with little or no accumulation in healthy organs 8 hours postinjection, and subsequently repressed tumor growth at low doses with high efficiency.

Here, the authors show that sequential treatment with long-acting slow-effective release ART and AAV9- based delivery of CRISPR-Cas9 results in undetectable levels of virus and integrated DNA in a subset of humanized HIV-1 infected mice. This proof-of-concept study suggests that HIV-1 elimination is possible.

n a step forward in the search for an HIV vaccine, Professor Denis Burton and colleagues from the Scripps Institute in La Jolla, California have manufactured an HIV vaccine that, with just one shot, induced six out of 12 monkeys to make antibodies that significantly delayed infection or (in two cases) even prevented it. The findings are published in the 15 January issue of Immunity.

In this work, the authors  assembled 12 transcription activator-like effector nucleases (TALENs) and five guide RNAs (gRNAs) for CRISPR system to knock out endogenous TCR expression. Using nuclease-expressing plasmid DNA, they achieved up to 19.9% and 12.2% knockout of TCR expression in primary T cells with CRISPR/Cas9 and TALENs, respectively. In contrast, delivery of TALEN mRNA by electroporation resulted in high viability and TCR knockout efficiencies of up to 78.8% for the TCR α chain and 81.2% for the β chain on day 6 after electroporation. 

In this study, a flexible aptamer-liposome-CRISPR/Cas9 chimera was designed to combine efficient delivery and increased flexibility. the chimera incorporated an RNA aptamer that specifically binds prostate cancer cells expressing the prostate-specific membrane antigen as a ligand. The approach described here provides a universal means of cell type–specific CRISPR/Cas9 delivery, which is a critical goal for the widespread therapeutic applicability of CRISPR/Cas9 or other nucleic acid drugs.

In this study, the authors show that the CRISPR/Cas9 system can restrict the JC polyomavirus (JCPyV) life cycle in cultured cells. They utilized CRISPR/Cas9 to target the noncoding control region and the late gene open reading frame of the JCPyV genome. They found significant inhibition of virus replication and viral protein expression in cells recipient of Cas9 together with JCPyV-specific single-guide RNA delivered prior to or after JCPyV infection.