Cell Reprogramming With CRISPR/Cas9 Based Transcriptional Regulation Systems

Shakirova, K. M.; Ovchinnikova, Viktoriia Y.; Дашинимаев, Э. Б.

doi:10.3389/fbioe.2020.00882

Cited by 34 publications

(17 citation statements)

References 147 publications

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“…As in vitro transcription/translation platforms are easily adaptable to high-throughput formats [18,19], the parallel testing of multiple Cas9 enzymes with altered properties could be achieved. In addition to testing mutant Cas9 variants, the system might also be useful to assess the properties of Cas9 fusion proteins for different purposes, e.g., base editing [20], epigenetic modification [21], or transcriptional activation/repression [22]. Once a robust Cas9 nuclease has been identified for a particular purpose, the BYL system can be used to produce RNPs for subsequent DNA-free delivery to the appropriate target cell/tissue.…”

Section: Discussionmentioning

confidence: 99%

Rapid production of SaCas9 in plant-based cell-free lysate for activity testing

Schiermeyer

Cerda‐Bennasser²,

Schmelter³

et al. 2021

Preprint

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Cas9 nucleases have become the most versatile tool for genome editing projects in a broad range of organisms. The recombinant production of Cas9 nuclease is desirable for in vitro activity assays or the preparation of ribonucleoproteins (RNPs) for DNA-free genome editing approaches. For the rapid production of Cas9, we explored the use of a recently established cell-free lysate from tobacco (Nicotiana tabacum L.) BY-2 cells. Using this system, the 130-kDa Cas9 nuclease from Staphylococcus aureus (SaCas9) was produced and subsequently purified via affinity chromatography. The purified apoenzyme was supplemented with ten different sgRNAs, and the nuclease activity was confirmed by the linearization of plasmid DNA containing cloned DNA target sequences.

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Section: Discussionmentioning

confidence: 99%

Rapid production of SaCas9 in plant-based cell-free lysate for activity testing

Schiermeyer

Cerda‐Bennasser²,

Schmelter³

et al. 2021

Preprint

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“…When directed to DNA regions using synthetic single guide RNAs (sgRNA), this increases the expression of endogenous genes of interest. Thus, the use of this technology with all its derivatives is of great interest for the fields of cell reprogramming and regenerative medicine [156,157].…”

Section: Crispr-cas9mentioning

confidence: 99%

RNA-Based Strategies for Cell Reprogramming toward Pluripotency

2022

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Cell therapy approaches to treat a wide range of pathologies have greatly benefited from cell reprogramming techniques that allow the conversion of a somatic cell into a pluripotent cell. Many technological developments have been made since the initial major discovery of this biological process. Recently reprogramming methods based on the use of RNA have emerged and seem very promising. Thus, in this review we will focus on presenting the interest of such methods for cell reprogramming but also how these RNA-based strategies can be extended to eventually lead to medical applications to improve healthspan and longevity.

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“…VP64 is a tetramer of VP16–a well-characterized herpes simplex virus transcription activator. It is among the first generation of CRISPRa systems and demonstrates a strong induction of activation in many experiments [ 63 ]. First-generation CRISPRa systems contain dCas9 fused with transactivator and sgRNA.…”

Section: Activation Of Defense Genes Through Crispr Activation (Crispra)mentioning

confidence: 99%

Control of Bacterial Diseases of Banana Using CRISPR/Cas-Based Gene Editing

Tripathi

Ntui

Tripathi

2022

IJMS

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Banana is an important staple food crop and a source of income for smallholder farmers in about 150 tropical and sub-tropical countries. Several bacterial diseases, such as banana Xanthomonas wilt (BXW), blood, and moko disease, cause substantial impacts on banana production. There is a vast yield gap in the production of bananas in regions where bacterial pathogens and several other pathogens and pests are present together in the same field. BXW disease caused by Xanthomonas campestris pv. musacearum is reported to be the most destructive banana disease in East Africa. The disease affects all the banana varieties grown in the region. Only the wild-type diploid banana, Musa balbisiana, is resistant to BXW disease. Developing disease-resistant varieties of bananas is one of the most effective strategies to manage diseases. Recent advances in CRISPR/Cas-based gene editing techniques can accelerate banana improvement. Some progress has been made to create resistance against bacterial pathogens using CRISPR/Cas9-mediated gene editing by knocking out the disease-causing susceptibility (S) genes or activating the expression of the plant defense genes. A synopsis of recent advancements and perspectives on the application of gene editing for the control of bacterial wilt diseases are presented in this article.

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Cell Reprogramming With CRISPR/Cas9 Based Transcriptional Regulation Systems

Cited by 34 publications

References 147 publications

Rapid production of SaCas9 in plant-based cell-free lysate for activity testing

Rapid production of SaCas9 in plant-based cell-free lysate for activity testing

RNA-Based Strategies for Cell Reprogramming toward Pluripotency

Control of Bacterial Diseases of Banana Using CRISPR/Cas-Based Gene Editing

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