scholarly journals SgRNA Expression of CRIPSR-Cas9 System Based on MiRNA Polycistrons as a Versatile Tool to Manipulate Multiple and Tissue-Specific Genome Editing

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Chen Xie ◽  
Yan-Lian Chen ◽  
Dong-Fang Wang ◽  
Yi-Lin Wang ◽  
Tian-Peng Zhang ◽  
...  
Keyword(s):  
Genome Editing ◽  
Tissue Specific ◽  
Versatile Tool

scholarly journals CRISPR-Cas12a (Cpf1): A Versatile Tool in the Plant Genome Editing Tool Box for Agricultural Advancement

2020 ◽  
Author(s):  
Anindya Bandyopadhyay ◽  
Nagesh Kancharla ◽  
vivek javalkote ◽  
santanu dasgupta ◽  
Thomas Brutnell
Keyword(s):  
Genome Editing ◽  
Genome Engineering ◽  
Temperature Stability ◽  
Double Strand Breaks ◽  
Plant Genome ◽  
Strand Breaks ◽  
Guide Rna ◽  
Versatile Tool ◽  
Type V ◽  
Global Population

Global population is predicted to approach 10 billion by 2050, an increase of over 2 billion from today. To meet the demands of growing, geographically and socio-economically diversified nations, we need to diversity and expand agricultural production. This expansion of agricultural productivity will need to occur under increasing biotic, and environmental constraints driven by climate change. Clustered regularly interspaced short palindromic repeats-site directed nucleases (CRISPR-SDN) and similar genome editing technologies will likely be key enablers to meet future agricultural needs. While the application of CRISPR-Cas9 mediated genome editing has led the way, the use of CRISPR-Cas12a is also increasing significantly for genome engineering of plants. The popularity of the CRISPR-Cas12a, the type V (class-II) system, is gaining momentum because of its versatility and simplified features. These include the use of a small guide RNA devoid of trans-activating crispr RNA (tracrRNA), targeting of T-rich regions of the genome where Cas9 is not suitable for use, RNA processing capability facilitating simpler multiplexing, and its ability to generate double strand breaks (DSB) with staggered ends. Many monocot and dicot species have been successfully edited using this Cas12a system and further research is ongoing to improve its efficiency in plants, including improving the temperature stability of the Cas12a enzyme, identifying new variants of Cas12a or synthetically producing Cas12a with flexible PAM sequences. In this review we provide a comparative survey of CRISPR-Cas12a and Cas9, and provide a perspective on applications of CRISPR-Cas12 in agriculture.

scholarly journals Short term but highly efficient Cas9 expression mediated by excisional system using adenovirus vector and Cre

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sayaka Nagamoto ◽  
Miyuki Agawa ◽  
Emi Tsuchitani ◽  
Kazunori Akimoto ◽  
Saki Kondo Matsushima ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Genome Editing ◽  
Adenovirus Vector ◽  
Virus Genome ◽  
Dna Viruses ◽  
Tissue Specific ◽  
B Virus ◽  
Tissue Specific Promoter ◽  
A Cell

AbstractGenome editing techniques such as CRISPR/Cas9 have both become common gene engineering technologies and have been applied to gene therapy. However, the problems of increasing the efficiency of genome editing and reducing off-target effects that induce double-stranded breaks at unexpected sites in the genome remain. In this study, we developed a novel Cas9 transduction system, Exci-Cas9, using an adenovirus vector (AdV). Cas9 was expressed on a circular molecule excised by the site-specific recombinase Cre and succeeded in shortening the expression period compared to AdV, which expresses the gene of interest for at least 6 months. As an example, we chose hepatitis B, which currently has more than 200 million carriers in the world and frequently progresses to liver cirrhosis or hepatocellular carcinoma. The efficiencies of hepatitis B virus genome disruption by Exci-Cas9 and Cas9 expression by AdV directly (Avec) were the same, about 80–90%. Furthermore, Exci-Cas9 enabled cell- or tissue-specific genome editing by expressing Cre from a cell- or tissue-specific promoter. We believe that Exci-Cas9 developed in this study is useful not only for resolving the persistent expression of Cas9, which has been a problem in genome editing, but also for eliminating long-term DNA viruses such as human papilloma virus.

scholarly journals Genome Editing as A Versatile Tool to Improve Horticultural Crop Qualities

2020 ◽  
Vol 6 (6) ◽  
pp. 372-384
Author(s):  
Yating Chen ◽  
Wenwen Mao ◽  
Ting Liu ◽  
Qianqian Feng ◽  
Li Li ◽  
...  
Keyword(s):  
Genome Editing ◽  
Horticultural Crop ◽  
Versatile Tool

scholarly journals Tissue Specific DNA Repair Outcomes Shape the Landscape of Genome Editing

2021 ◽  
Vol 12 ◽  
Author(s):  
Mathilde Meyenberg ◽  
Joana Ferreira da Silva ◽  
Joanna I. Loizou
Keyword(s):  
Dna Repair ◽  
Genome Editing ◽  
Genetic Diseases ◽  
Dna Double Strand Breaks ◽  
Dna Breaks ◽  
Dna Damage And Repair ◽  
Tissue Specific ◽  
Precise Control ◽  
Repair Processes ◽  
Recent Developments

The use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 has moved from bench to bedside in less than 10years, realising the vision of correcting disease through genome editing. The accuracy and safety of this approach relies on the precise control of DNA damage and repair processes to achieve the desired editing outcomes. Strategies for modulating pathway choice for repairing CRISPR-mediated DNA double-strand breaks (DSBs) have advanced the genome editing field. However, the promise of correcting genetic diseases with CRISPR-Cas9 based therapies is restrained by a lack of insight into controlling desired editing outcomes in cells of different tissue origin. Here, we review recent developments and urge for a greater understanding of tissue specific DNA repair processes of CRISPR-induced DNA breaks. We propose that integrated mapping of tissue specific DNA repair processes will fundamentally empower the implementation of precise and safe genome editing therapies for a larger variety of diseases.

scholarly journals CRISPR-Cas Genome editing system: a versatile tool for developing disease resistant crops

Plant Stress ◽  
2022 ◽  
pp. 100056
Author(s):  
Ashwini Talakayala ◽  
Srinivas Ankanagari ◽  
Mallikarjuna Garladinne
Keyword(s):  
Genome Editing ◽  
System A ◽  
Disease Resistant ◽  
Versatile Tool

scholarly journals zGrad is a nanobody-based degron system that inactivates proteins in zebrafish

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Naoya Yamaguchi ◽  
Tugba Colak-Champollion ◽  
Holger Knaut
Keyword(s):  
Rna Interference ◽  
Protein Function ◽  
Fusion Proteins ◽  
Gene Activity ◽  
Modern Biology ◽  
Tissue Specific ◽  
Inducible Promoters ◽  
Protein Depletion ◽  
Protein Inactivation ◽  
Versatile Tool

The analysis of protein function is essential to modern biology. While protein function has mostly been studied through gene or RNA interference, more recent approaches to degrade proteins directly have been developed. Here, we adapted the anti-GFP nanobody-based system deGradFP from flies to zebrafish. We named this system zGrad and show that zGrad efficiently degrades transmembrane, cytosolic and nuclear GFP-tagged proteins in zebrafish in an inducible and reversible manner. Using tissue-specific and inducible promoters in combination with functional GFP-fusion proteins, we demonstrate that zGrad can inactivate transmembrane and cytosolic proteins globally, locally and temporally with different consequences. Global protein depletion results in phenotypes similar to loss of gene activity, while local and temporal protein inactivation yields more restricted and novel phenotypes. Thus, zGrad is a versatile tool to study the spatial and temporal requirement of proteins in zebrafish.

scholarly journals Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9

Development ◽  
2014 ◽  
Vol 141 (21) ◽  
pp. 4115-4120 ◽  
Author(s):  
A. Stolfi ◽  
S. Gandhi ◽  
F. Salek ◽  
L. Christiaen
Keyword(s):  
Genome Editing ◽  
Tissue Specific
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