Prime editing in plants and mammalian cells: Mechanism, achievements, limitations, and future prospects

Hillary, V. Edwin; Ceasar, Stanislaus Antony

doi:10.1002/bies.202200032

Cited by 30 publications

(17 citation statements)

References 79 publications

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“…Like Cas9, Cas12 protein was also considered as a sole member of the CRISPR family for genome editing. But in most circumstances, Cas12 was deemed superior to Cas9 protein because Cas12 protein generates staggered DSBs and promotes HDR repair mechanism instead of both NHEJ & HDR [ 67 ]. Cas12 system also overcomes disadvantages associated with diagnostic strategies.…”

Section: Cas12 Proteinmentioning

confidence: 99%

A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering

2022

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The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (CRISPR/Cas) system has altered life science research offering enormous options in manipulating, detecting, imaging, and annotating specific DNA or RNA sequences of diverse organisms. This system incorporates fragments of foreign DNA (spacers) into CRISPR cassettes, which are further transcribed into the CRISPR arrays and then processed to make guide RNA (gRNA). The CRISPR arrays are genes that encode Cas proteins. Cas proteins provide the enzymatic machinery required for acquiring new spacers targeting invading elements. Due to programmable sequence specificity, numerous Cas proteins such as Cas9, Cas12, Cas13, and Cas14 have been exploited to develop new tools for genome engineering. Cas variants stimulated genetic research and propelled the CRISPR/Cas tool for manipulating and editing nucleic acid sequences of living cells of diverse organisms. This review aims to provide detail on two classes (class 1 and 2) of the CRISPR/Cas system, and the mechanisms of all Cas proteins, including Cas12, Cas13, and Cas14 discovered so far. In addition, we also discuss the pros and cons and recent applications of various Cas proteins in diverse fields, including those used to detect viruses like severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). This review enables the researcher to gain knowledge on various Cas proteins and their applications, which have the potential to be used in next-generation precise genome engineering.

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Section: Cas12 Proteinmentioning

confidence: 99%

A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering

2022

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Section: Challenges Of Pementioning

confidence: 99%

“…PE3‐induced alteration restored normal mRNA splicing of β‐globin, and the corrected mutation had the same phenotype as the wild‐type IVS‐II‐654. Furthermore, analysis of the off‐target effects of nick sgRNA and pegRNA demonstrated PE system 3's genomic safety (Hillary & Ceasar, 2022). Another study used PE3 to efficiently induce the HBB mutation of β‐thalassemia in the HUDEP‐2 erythroid progenitor cell line.…”

Section: Potential Of Pe In β‐Thalassemiamentioning

confidence: 99%

“…In conclusion, despite the technical challenges and limitations, it is clear that PE can play a pivotal role in the near future as a genome editing technology (Hillary & Ceasar, 2022; Scholefield & Harrison, 2021). However, future research is required on in‐depth understanding of the PE design, extension of the editing window, and optimization of factors affecting editing efficiency to overcome the above‐mentioned key limitations of this technology.…”

Section: Challenges Of Pementioning

confidence: 99%

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Prime editing: A potential treatment option for β‐thalassemia

Arif

Farooq

Ahmad

et al. 2022

Cell Biology International

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The potential to therapeutically alter the genome is one of the remarkable scientific developments in recent years. Genome editing technologies have provided an opportunity to precisely alter genomic sequence(s) in eukaryotic cells as a treatment option for various genetic disorders. These technologies allow the correction of harmful mutations in patients by precise nucleotide editing. Genome editing technologies such as CRISPR (clustered regularly interspaced short palindromic repeat) and base editors have greatly contributed to the practical applications of gene editing. However, these technologies have certain limitations, including imperfect editing, undesirable mutations, off‐target effects, and lack of potential to simultaneously edit multiple loci. Recently, prime editing (PE) has emerged as a new gene editing technology with the potential to overcome the above‐mentioned limitations. Interestingly, PE not only has higher specificity but also does not require double‐strand breaks. In addition, a minimum possibility of potential off‐target mutant sites makes PE a preferred choice for therapeutic gene editing. Furthermore, PE has the potential to introduce insertion and deletions of all 12 single‐base mutations at target sequences. Considering its potential, PE has been applied as a treatment option for genetic diseases including hemoglobinopathies. β‐Thalassemia, for example, one of the most significant blood disorders characterized by reduced levels of functional hemoglobin, could potentially be treated using PE. Therapeutic reactivation of the γ‐globin gene in adult β‐thalassemia patients through PE technology is considered a promising therapeutic strategy. The current review aims to briefly discuss the genome editing strategies and potential applications of PE for the treatment of β‐thalassemia. In addition, the review will also focus on challenges associated with the use of PE.

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“…The OsLsi gene family has been reported to be involved in Si xylem loading in rice roots, which is required for efficient Si transport from roots to stems. CRISPR-Cas-based inactivation of this gene (OsLsi) resulted in reduced Si uptake in xylem sap and also reduced As accumulation in rice [33]. Similarly, CRISPR/Cas9 has been used to knock down OsNramp5 and OsLCT1 to reduce cadmium (Cd) accumulation [104] and inactivate the low cesium (Cs) K+ transporter OsHAK1 in rice plants [94].…”

Section: Crispr/cas Genome Editingmentioning

confidence: 99%

Recent Developments in Rice Molecular Breeding for Tolerance to Heavy Metal Toxicity

et al. 2023

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Heavy metal toxicity generally refers to the negative impact on the environment, humans, and other living organisms caused by exposure to heavy metals (HMs). Heavy metal poisoning is the accumulation of HMs in the soft tissues of organisms in a toxic amount. HMs bind to certain cells and prevent organs from functioning. Symptoms of HM poisoning can be life-threatening and not only cause irreversible damage to humans and other organisms; but also significantly reduce agricultural yield. Symptoms and physical examination findings associated with HM poisoning vary depending on the metal accumulated. Many HMs, such as zinc, copper, chromium, iron, and manganese, are present at extremely low levels but are essential for the functioning of plants. However, if these metals accumulate in the plants in sufficient concentrations to cause poisoning, serious damage can occur. Rice is consumed around the world as a staple food and incidents of HM pollution often occur in rice-growing areas. In many rice-producing countries, cadmium (Cd), arsenic (As), and lead (Pb) have been recognized as commonly widespread HMs contaminating rice fields worldwide. In addition to mining and irrigation activities, the use of fertilizers and pesticides has also contributed significantly to HM contamination of rice-growing soils around the world. A number of QTLs associated with HM stress signals from various intermediary molecules have been reported to activate various transcription factors (TFs). Some antioxidant enzymes have been studied which contribute to the scavenging of reactive oxygen species, ultimately leading to stress tolerance in rice. Genome engineering and advanced editing techniques have been successfully applied to rice to improve metal tolerance and reduce HM accumulation in grains. In this review article, recent developments and progress in the molecular science for the induction of HM stress tolerance, including reduced metal uptake, compartmentalized transportation, gene-regulated signaling, and reduced accumulation or diversion of HM particles to plant parts other than grains, are discussed in detail, with particular emphasis on rice.

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Prime editing in plants and mammalian cells: Mechanism, achievements, limitations, and future prospects

Cited by 30 publications

References 79 publications

A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering

A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering

Prime editing: A potential treatment option for β‐thalassemia

Recent Developments in Rice Molecular Breeding for Tolerance to Heavy Metal Toxicity

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