Adenoviral vector DNA for accurate genome editing with engineered nucleases

SummaryTargeted genome editing with designer nucleases, such as zinc finger nucleases, TALE nucleases, and CRISPR-Cas nucleases, has heralded a new era in gene therapy. Genetic disorders, which have not been amenable to conventional gene-addition-type gene therapy approaches, such as disorders with dominant inheritance or diseases caused by mutations in tightly regulated genes, can now be treated by precise genome surgery. Moreover, engineered nucleases enable novel genetic interventions to fight infectious diseases or to improve cancer immunotherapies. Here, we review the development of the different classes of programmable nucleases, discuss the challenges and improvements in translating gene editing into clinical use, and give an outlook on what applications can expect to enter the clinic in the near future.

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Oligonucleotide-Mediated Genome Editing Provides Precision and Function to Engineered Nucleases and Antibiotics in Plants

PLANT PHYSIOLOGY ◽

10.1104/pp.15.01696 ◽

2016 ◽

Vol 170 (4) ◽

pp. 1917-1928 ◽

Cited By ~ 84

Author(s):

Noel J. Sauer ◽

Javier Narváez-Vásquez ◽

Jerry Mozoruk ◽

Ryan B. Miller ◽

Zachary J. Warburg ◽

...

Keyword(s):

Genome Editing ◽

Engineered Nucleases ◽

And Function

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Genome Editing with Engineered Nucleases in Economically Important Animals and Plants: State of the Art in the Research Pipeline

Current Issues in Molecular Biology ◽

10.21775/cimb.021.041 ◽

2017 ◽

Cited By ~ 4

Keyword(s):

Genome Editing ◽

State Of The Art ◽

Engineered Nucleases

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Comparison of T7E1 and Surveyor Mismatch Cleavage Assays to Detect Mutations Triggered by Engineered Nucleases

G3 Genes|Genome|Genetics ◽

10.1534/g3.114.015834 ◽

2015 ◽

Vol 5 (3) ◽

pp. 407-415 ◽

Cited By ~ 156

Author(s):

Léna Vouillot ◽

Aurore Thélie ◽

Nicolas Pollet

Keyword(s):

Genome Editing ◽

Mutation Detection ◽

Point Mutations ◽

Targeted Mutagenesis ◽

Single Nucleotide ◽

Heteroduplex Dna ◽

Hit Rate ◽

Engineered Nucleases ◽

Mismatch Cleavage ◽

Surveyor Nuclease

Abstract Genome editing using engineered nucleases is used for targeted mutagenesis. But because genome editing does not target all loci with similar efficiencies, the mutation hit-rate at a given locus needs to be evaluated. The analysis of mutants obtained using engineered nucleases requires specific methods for mutation detection, and the enzyme mismatch cleavage method is used commonly for this purpose. This method uses enzymes that cleave heteroduplex DNA at mismatches and extrahelical loops formed by single or multiple nucleotides. Bacteriophage resolvases and single-stranded nucleases are used commonly in the assay but have not been compared side-by-side on mutations obtained by engineered nucleases. We present the first comparison of the sensitivity of T7E1 and Surveyor EMC assays on deletions and point mutations obtained by zinc finger nuclease targeting in frog embryos. We report the mutation detection limits and efficiencies of T7E1 and Surveyor. In addition, we find that T7E1 outperforms the Surveyor nuclease in terms of sensitivity with deletion substrates, whereas Surveyor is better for detecting single nucleotide changes. We conclude that T7E1 is the preferred enzyme to scan mutations triggered by engineered nucleases.

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The position of the target site for engineered nucleases improves the aberrant mRNA clearance in in vivo genome editing

Scientific Reports ◽

10.1038/s41598-020-61154-4 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 2

Author(s):

Jae Hoon Lee ◽

Sungsook Yu ◽

Tae Wook Nam ◽

Jae-il Roh ◽

Young Jin ◽

...

Keyword(s):

Genome Editing ◽

Target Site ◽

Engineered Nucleases

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A therapeutic genome editing primer for cardiologist

Cardiogenetics ◽

10.4081/cardiogenetics.2018.7262 ◽

2018 ◽

Vol 8 (1) ◽

Author(s):

Martina Caiazza ◽

Daniele Masarone ◽

Giuseppe Limongelli

Keyword(s):

Lipid Metabolism ◽

Animal Models ◽

Genome Editing ◽

Genome Engineering ◽

Living Organism ◽

Cardiovascular Disorder ◽

Cardiovascular Disorders ◽

New Class ◽

Engineered Nucleases

Genome editing, or genome engineering is a type of genetic engineering in which DNA is inserted, deleted or replaced in the genome of a living organism using engineered nucleases, or molecular scissors. Genome editing is being rapidly adopted into all fields of biomedical research, including the cardiovascular field, where it has facilitated a greater understanding of lipid metabolism, electrophysiology, cardiomyopathies, and other cardiovascular disorders, has helped to create a wider variety of cellular and animal models, and has opened the door to a new class of therapies. In this review, we discuss the applications of in vivo genome-editing therapies for cardiovascular disorder.

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