Tools for Efficient Genome Editing; ZFN, TALEN, and CRISPR

Shamshirgaran, Yasaman; Li, Jun; Sümer, Hüseyin; Verma, Paul J.; Taheri‐Ghahfarokhi, Amir

doi:10.1007/978-1-0716-2301-5_2

Cited by 35 publications

(19 citation statements)

References 83 publications

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“…The application of PARS should be extended to other gene editing methods, including zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) ( Zhang et al., 2017 ; Shamshirgaran et al., 2022 ). As PAR1 is conserved among different plants tested ( Figure 6 ), we propose that the PARS system has the potential to be widely used in the gene editing of various plant species using multiple gene editing systems.…”

Section: Discussionmentioning

confidence: 99%

A simple and efficient strategy to produce transgene-free gene edited plants in one generation using paraquat resistant 1 as a selection marker

et al. 2023

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IntroductionDNA integration is a key factor limiting the marketing of CRISPR/Cas9-mediated gene edited crops. Several strategies have been established to obtain transgene-free gene edited plants; however, these strategies are usually time-consuming, technically difficult, providing low mutagenesis efficiency, and/or including a narrow host range.MethodTo overcome such issues, we established a paraquat resistant 1 (PAR1)-based positive screening (PARS) strategy, which achieved efficient screening of transgene-free gene edited plants. With PARS, the screening efficiency of mutant increased by 2.81-fold on average, and approximately 10% of T1 plants selected via PARS were transgene-free.ResultsMoreover, heritable transgene-free mutations at target loci were identified in the T1 generation. Based on the previous reports and our data, we know that paraquat is toxic to all green plants, PAR1 is conserved among all plant species tested, and the transient expression of Cas9 editor can produce transgene-free gene edited plants.DiscussionThus, we assume that the PARS strategy established here has the potential to be widely used to screen transgene-free mutants in various crops using diverse CRISPR/Cas9 delivery approaches.

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Section: Discussionmentioning

confidence: 99%

A simple and efficient strategy to produce transgene-free gene edited plants in one generation using paraquat resistant 1 as a selection marker

et al. 2023

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“…Although the CRISPR/Cas system is the most widely used genome editing strategy, TALENs are more effective in HDR repair [ 51 , 52 ]. However, CRISPR/Cas9 technology is more efficient, powerful, and flexible [ 53 , 54 ]. Unfortunately, we have not been able to obtain corrected clones through TALENs-mediated gene editing.…”

Section: Discussionmentioning

confidence: 99%

High-Efficiency CRISPR/Cas9-Mediated Correction of a Homozygous Mutation in Achromatopsia-Patient-Derived iPSCs

Siles

Gaudó

Pomares

2023

IJMS

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Achromatopsia is an autosomal recessive disorder, in which cone photoreceptors undergo progressive degeneration, causing color blindness and poor visual acuity, among other significant eye affectations. It belongs to a group of inherited retinal dystrophies thatcurrently have no treatment. Although functional improvements have been reported in several ongoing gene therapy studies, more efforts and research should be carried out to enhance their clinical application. In recent years, genome editing has arisen as one of the most promising tools for personalized medicine. In this study, we aimed to correct a homozygous PDE6C pathogenic variant in hiPSCs derived from a patient affected by achromatopsia through CRISPR/Cas9 and TALENs technologies. Here, we demonstrate high efficiency in gene editing by CRISPR/Cas9 but not with TALENs approximation. Despite a few of the edited clones displaying heterozygous on-target defects, the proportion of corrected clones with a potentially restored wild-type PDE6C protein was more than half of the total clones analyzed. In addition, none of them presented off-target aberrations. These results significantly contribute to advances in single-nucleotide gene editing and the development of future strategies for the treatment of achromatopsia.

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“…Genome editing tools hold great promise in treating genetic and non-genetic diseases. Early studies utilize zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) for genome editing ( Urnov et al, 2010 ; Joung and Sander, 2013 ; Shamshirgaran et al, 2022 ). ZFNs and TALENs depend on protein engineering of DNA-binding domains to recognize and edit specific DNA sequences.…”

Section: Introductionmentioning

confidence: 99%

Off-target effects in CRISPR/Cas9 gene editing

Guo

Gao

et al. 2023

Front. Bioeng. Biotechnol.

152

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Gene editing stands for the methods to precisely make changes to a specific nucleic acid sequence. With the recent development of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, gene editing has become efficient, convenient and programmable, leading to promising translational studies and clinical trials for both genetic and non-genetic diseases. A major concern in the applications of the CRISPR/Cas9 system is about its off-target effects, namely the deposition of unexpected, unwanted, or even adverse alterations to the genome. To date, many methods have been developed to nominate or detect the off-target sites of CRISPR/Cas9, which laid the basis for the successful upgrades of CRISPR/Cas9 derivatives with enhanced precision. In this review, we summarize these technological advancements and discuss about the current challenges in the management of off-target effects for future gene therapy.

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Tools for Efficient Genome Editing; ZFN, TALEN, and CRISPR

Cited by 35 publications

References 83 publications

A simple and efficient strategy to produce transgene-free gene edited plants in one generation using paraquat resistant 1 as a selection marker

A simple and efficient strategy to produce transgene-free gene edited plants in one generation using paraquat resistant 1 as a selection marker

High-Efficiency CRISPR/Cas9-Mediated Correction of a Homozygous Mutation in Achromatopsia-Patient-Derived iPSCs

Off-target effects in CRISPR/Cas9 gene editing

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