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Dear Colleagues,

Some genetic diseases affect only certain tissues, whereas others affect every cell in the body. For many of these diseases, intravenous delivery is favored to reach distant diseased tissues. While systemic delivery has some ability to do this, it also exposes many more off-target cells and tissues to the therapy. This increases the risk of toxic and immunologic side effects. Efficacy is frequently examined in gene therapy. Side-effects and toxicity are frequently ignored.

This Issue focuses on strategies to target vectors to the cells in need of therapy while also detargeting them from off-target sites. These strategies may include screening different vectors or capsids, modifying vectors with cell-targeting ligands, and post-entry strategies to control expression of transgenes in on-target tissues. Articles are encouraged that successfully track gene delivery by measuring transgene expression, but that also track where all of the failed vectors go by tracking vector genomes. Articles that examine vector efficacy in combination with measurements of gene therapy side effects are most encouraged.

Prof. Michael A. Barry
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Genes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

Gene therapy
Gene-based vaccines
On-target targeting
Off-target targeting
Immunology
Anti-cancer vaccines
Viral vectors
Adenoviruses

Published Papers (2 papers)

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Research

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17 pages, 4385 KiB

Open AccessArticle

Unlocking loxP to Track Genome Editing In Vivo

by William A. C. Gendron, Jeffrey D. Rubin, Michael J. Hansen, Rebecca A. Nace, Brandon W. Simone, Stephen C. Ekker and Michael A. Barry

Genes 2021, 12(8), 1204; https://0-doi-org.brum.beds.ac.uk/10.3390/genes12081204 - 03 Aug 2021

Cited by 1 | Viewed by 4225

Abstract

The development of CRISPR-associated proteins, such as Cas9, has led to increased accessibility and ease of use in genome editing. However, additional tools are needed to quantify and identify successful genome editing events in living animals. We developed a method to rapidly quantify and monitor gene editing activity non-invasively in living animals that also facilitates confocal microscopy and nucleotide level analyses. Here we report a new CRISPR “fingerprinting” approach to activating luciferase and fluorescent proteins in mice as a function of gene editing. This system is based on experience with our prior cre recombinase (cre)-detector system and is designed for Cas editors able to target loxP including gRNAs for SaCas9 and ErCas12a. These CRISPRs cut specifically within loxP, an approach that is a departure from previous gene editing in vivo activity detection techniques that targeted adjacent stop sequences. In this sensor paradigm, CRISPR activity was monitored non-invasively in living cre reporter mice (FVB.129S6(B6)-Gt(ROSA)26Sortm1(Luc)Kael/J and Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J, which will be referred to as LSL-luciferase and mT/mG throughout the paper) after intramuscular or intravenous hydrodynamic plasmid injections, demonstrating utility in two diverse organ systems. The same genome-editing event was examined at the cellular level in specific tissues by confocal microscopy to determine the identity and frequency of successfully genome-edited cells. Further, SaCas9 induced targeted editing at efficiencies that were comparable to cre, demonstrating high effective delivery and activity in a whole animal. This work establishes genome editing tools and models to track CRISPR editing in vivo non-invasively and to fingerprint the identity of targeted cells. This approach also enables similar utility for any of the thousands of previously generated loxP animal models. Full article

(This article belongs to the Special Issue Balancing On Target and Off Target Delivery during Systemic Gene Therapy)

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Graphical abstract

Review

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32 pages, 1403 KiB

Open AccessReview

Delivery Approaches for Therapeutic Genome Editing and Challenges

by Ilayda Ates, Tanner Rathbone, Callie Stuart, P. Hudson Bridges and Renee N. Cottle

Genes 2020, 11(10), 1113; https://0-doi-org.brum.beds.ac.uk/10.3390/genes11101113 - 23 Sep 2020

Cited by 35 | Viewed by 5941

Abstract

Impressive therapeutic advances have been possible through the advent of zinc-finger nucleases and transcription activator-like effector nucleases. However, discovery of the more efficient and highly tailorable clustered regularly interspaced short palindromic repeats (CRISPR) and associated proteins (Cas9) has provided unprecedented gene-editing capabilities for treatment of various inherited and acquired diseases. Despite recent clinical trials, a major barrier for therapeutic gene editing is the absence of safe and effective methods for local and systemic delivery of gene-editing reagents. In this review, we elaborate on the challenges and provide practical considerations for improving gene editing. Specifically, we highlight issues associated with delivery of gene-editing tools into clinically relevant cells. Full article

(This article belongs to the Special Issue Balancing On Target and Off Target Delivery during Systemic Gene Therapy)

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