A Review on Advanced CRISPR-Based Genome-Editing Tools: Base Editing and Prime Editing

Sichani, Ali Saber; Ranjbar, Maryam; Baneshi, Maryam; Zadeh, Farid Torabi; Fallahi, Jafar

doi:10.1007/s12033-022-00639-1

Cited by 22 publications

(11 citation statements)

References 84 publications

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“…To improve the efficiency of HDR-mediated editing, base editors that enable precise base editing have been developed. As some of the SpCas9 variant enzymes can recognize unconventional PAMs (e.g., XCas9 and SPCas9-NG), an increasing number of variant enzymes are being used in base editing [ 104 , 105 , 106 ]. In 2019, Xu et al developed three high-fidelity SpCas9 variants, eSpCas9, SpCas9-HF2, and HypaCas9, designed to act as C-T base editors together with PmCDA1 (pBEs).…”

Section: Strategies and Methods For Optimizing Crispr/cas9 Gene-editi...mentioning

confidence: 99%

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Zhou

Luan

Liu

et al. 2023

Plants

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Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature and is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation of transgene-free genome-edited plants in a short period and has the advantages of simplicity, high efficiency, high specificity, and low production costs, which greatly facilitate the study of gene functions. In plant molecular breeding, the gene-editing efficiency of the CRISPR-Cas9 system has proven to be a key step in influencing the effectiveness of molecular breeding, with improvements in gene-editing efficiency recently becoming a focus of reported scientific research. This review details strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding, including Cas9 variant enzyme engineering, the effect of multiple promoter driven Cas9, and gRNA efficient optimization and expression strategies. It also briefly introduces the optimization strategies of the CRISPR/Cas12a system and the application of BE and PE precision editing. These strategies are beneficial for the further development and optimization of gene editing systems in the field of plant molecular breeding.

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Section: Strategies and Methods For Optimizing Crispr/cas9 Gene-editi...mentioning

confidence: 99%

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Zhou

Luan

Liu

et al. 2023

Plants

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show abstract

“…insertion of a stretch of DNA or base pair modification) will trigger homology-directed repair (HDR), resulting in gene knock in or base editing . Practical considerations of CRISPR/Cas9-mediated gene knock-in and base editing will not be addressed in detail but are expertly discussed in refs − .…”

Section: Orthogonal Validationmentioning

confidence: 99%

Comprehensive Overview of Bottom-Up Proteomics Using Mass Spectrometry

Jiang,

Rex,

Schuster

et al. 2024

ACS Meas. Sci. Au

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Proteomics is the large scale study of protein structure and function from biological systems through protein identification and quantification. “Shotgun proteomics” or “bottom-up proteomics” is the prevailing strategy, in which proteins are hydrolyzed into peptides that are analyzed by mass spectrometry. Proteomics studies can be applied to diverse studies ranging from simple protein identification to studies of proteoforms, protein-protein interactions, protein structural alterations, absolute and relative protein quantification, post-translational modifications, and protein stability. To enable this range of different experiments, there are diverse strategies for proteome analysis. The nuances of how proteomic workflows differ may be challenging to understand for new practitioners. Here, we provide a comprehensive overview of different proteomics methods. We cover from biochemistry basics and protein extraction to biological interpretation and orthogonal validation. We expect this Review will serve as a handbook for researchers who are new to the field of bottom-up proteomics.

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“…Using bioinformatics tools, researchers can identify and avoid gRNAs with off-target sites in the genome. Base editing and prime editing: instead of creating double-strand breaks, base editors and prime editors can perform precise single-base changes without introducing DSBs, reducing the potential for off-target effects [ 70 , 71 , 72 ]. Targeted liposome delivery: by modifying liposomes with ligands that specifically recognize cell surface markers or receptors unique to the target cells, liposomes can be directed to the desired cells or tissues.…”

Section: Challenges For Liposome-based Crispr Genome Editingmentioning

confidence: 99%

Liposome-Based Carriers for CRISPR Genome Editing

Yin

Harmancey

McPherson

et al. 2023

IJMS

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The CRISPR-based genome editing technology, known as clustered regularly interspaced short palindromic repeats (CRISPR), has sparked renewed interest in gene therapy. This interest is accompanied by the development of single-guide RNAs (sgRNAs), which enable the introduction of desired genetic modifications at the targeted site when used alongside the CRISPR components. However, the efficient delivery of CRISPR/Cas remains a challenge. Successful gene editing relies on the development of a delivery strategy that can effectively deliver the CRISPR cargo to the target site. To overcome this obstacle, researchers have extensively explored non-viral, viral, and physical methods for targeted delivery of CRISPR/Cas9 and a guide RNA (gRNA) into cells and tissues. Among those methods, liposomes offer a promising approach to enhance the delivery of CRISPR/Cas and gRNA. Liposomes facilitate endosomal escape and leverage various stimuli such as light, pH, ultrasound, and environmental cues to provide both spatial and temporal control of cargo release. Thus, the combination of the CRISPR-based system with liposome delivery technology enables precise and efficient genetic modifications in cells and tissues. This approach has numerous applications in basic research, biotechnology, and therapeutic interventions. For instance, it can be employed to correct genetic mutations associated with inherited diseases and other disorders or to modify immune cells to enhance their disease-fighting capabilities. In summary, liposome-based CRISPR genome editing provides a valuable tool for achieving precise and efficient genetic modifications. This review discusses future directions and opportunities to further advance this rapidly evolving field.

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A Review on Advanced CRISPR-Based Genome-Editing Tools: Base Editing and Prime Editing

Cited by 22 publications

References 84 publications

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Comprehensive Overview of Bottom-Up Proteomics Using Mass Spectrometry

Liposome-Based Carriers for CRISPR Genome Editing

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