New Bicistronic TALENs Greatly Improve Genome Editing

Martín-Fernández, José María; Fleischer, Aarne; Vallejo-Díez, Sara; Palomino, Esther; Sánchez‐Gilabert, Almudena; Ruiz, Raúl; Bejarano, Yazmine; Llinàs, Pere; Gayà, Antoni; Bachiller, Daniel

doi:10.1002/cpsc.104

Cited by 7 publications

(4 citation statements)

References 47 publications

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“…This improvement could help each TALEN monomer to transcribe from the same reading frame in order to increase the gene editing efficacy. ( Mariano et al, 2014 ; Martín-Fernández et al, 2020 ). Additionally, Zhang et al (2017) used deciphered TALEs for 5-hydroxymethylcytosine and 5-methylcytosine to achieve methylation-dependent genome editing and gene activation in vivo .…”

Section: Nuclease-based Genome Engineering Technologiesmentioning

confidence: 99%

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Mbakam²,

Song³

et al. 2022

Front. Genome Ed.

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Advancements in genome editing make possible to exploit the functions of enzymes for efficient DNA modifications with tremendous potential to treat human genetic diseases. Several nuclease genome editing strategies including Meganucleases (MNs), Zinc Finger Nucleases (ZFNs), Transcription Activator-like Effector Nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated proteins (CRISPR-Cas) have been developed for the correction of genetic mutations. CRISPR-Cas has further been engineered to create nickase genome editing tools including Base editors and Prime editors with much precision and efficacy. In this review, we summarized recent improvements in nuclease and nickase genome editing approaches for the treatment of genetic diseases. We also highlighted some limitations for the translation of these approaches into clinical applications.

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Section: Nuclease-based Genome Engineering Technologiesmentioning

confidence: 99%

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Mbakam²,

Song³

et al. 2022

Front. Genome Ed.

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“…Using a bicistronic TALEN construct which transcribes two proteins from the same plasmid, about 15% higher cleavage activity of TALENs was achieved as compared to their separate expression as two monomers in cotransfection [ 58 ]. Furthermore, bicistronic TALENs enable real-time monitoring of transfection efficiency and rapid enrichment selection of nuclease-containing cells [ 61 ].…”

Section: Talensmentioning

confidence: 99%

Comparison of the Feasibility, Efficiency, and Safety of Genome Editing Technologies

Castro

Bjelic

Malhotra

et al. 2021

IJMS

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Recent advances in programmable nucleases including meganucleases (MNs), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats-Cas (CRISPR-Cas) have propelled genome editing from explorative research to clinical and industrial settings. Each technology, however, features distinct modes of action that unevenly impact their applicability across the entire genome and are often tested under significantly different conditions. While CRISPR-Cas is currently leading the field due to its versatility, quick adoption, and high degree of support, it is not without limitations. Currently, no technology can be regarded as ideal or even applicable to every case as the context dictates the best approach for genetic modification within a target organism. In this review, we implement a four-pillar framework (context, feasibility, efficiency, and safety) to assess the main genome editing platforms, as a basis for rational decision-making by an expanding base of users, regulators, and consumers. Beyond carefully considering their specific use case with the assessment framework proposed here, we urge stakeholders interested in genome editing to independently validate the parameters of their chosen platform prior to commitment. Furthermore, safety across all applications, particularly in clinical settings, is a paramount consideration and comprehensive off-target detection strategies should be incorporated within workflows to address this. Often neglected aspects such as immunogenicity and the inadvertent selection of mutants deficient for DNA repair pathways must also be considered.

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“…To simplify the selection process, we thought to exploit the use of a magnetic-activated cell sorting system (Duda et al, 2014;Martin-Fernandez et al, 2020) to sort CRISPR genetically modified cells. Here, we describe a new method called "magnetic-activated genomeedited cells sorting" assay or MAGECS.…”

Section: Introductionmentioning

confidence: 99%

Fast but not furious: a streamlined selection method for genome-edited cells

et al. 2021

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In the last decade, transcription activator-like effector nucleases and CRISPR-based genome engineering have revolutionized our approach to biology. Because of their high efficiency and ease of use, the development of custom knock-out and knock-in animal or cell models is now within reach for almost every laboratory. Nonetheless, the generation of genetically modified cells often requires a selection step, usually achieved by antibiotics or fluorescent markers. The choice of the selection marker is based on the available laboratory resources, such as cell types, and parameters such as time and cost should also be taken into consideration. Here, we present a new and fast strategy called magnetic-activated genome-edited cell sorting, to select genetically modified cells based on the ability to magnetically sort surface antigens (i.e., tCD19) present in Cas9-positive cells. By using magnetic-activated genome-edited cell sorting, we successfully generated and isolated genetically modified human-induced pluripotent stem cells, primary human fibroblasts, SH-SY5Y neuroblast-like cells, HaCaT and HEK 293T cells. Our strategy expands the genome editing toolbox by offering a fast, cheap, and an easy to use alternative to the available selection methods.

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New Bicistronic TALENs Greatly Improve Genome Editing

Cited by 7 publications

References 47 publications

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Comparison of the Feasibility, Efficiency, and Safety of Genome Editing Technologies

Fast but not furious: a streamlined selection method for genome-edited cells

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