scholarly journals Current Capabilities, Precision, and Risks of Genome Editing Techniques in Agricultural Systems

2020 ◽  
Author(s):  
Morgan Carter
Keyword(s):  
Best Practices ◽  
Genome Editing ◽  
Target Selection ◽  
Agricultural Systems ◽  
Molecular Tools ◽  
A Genome ◽  
Pros And Cons ◽  
Regulatory Measures ◽  
Wide Testing ◽  
Careful Design

We are in a new chapter of crop and livestock improvement with the emergence of genome editing. This latest generation of molecular tools can be used to make targeted changes in a genome including insertions, deletions, and mutations. With new advances comes new risks for unintended changes and impacts, thus the need for appropriate risk assessment for product development and to inform regulatory measures. Though CRISPR/Cas has arisen as the predominant technology, there are multiple types of genome editing tools each with pros and cons depending on the organism and desired outcome. Furthermore, each editing tool differs in specificity as they may edit non-intended sites, referred to as off-target edits. The consensus of the agricultural editing community is to avoid off-target editing through design and detection, instead of determining whether off-target editing in each case is detrimental. The design of a targeting component, the tool chosen, and the identification of the edit(s) made are the critical factors in avoiding off-target edits and confirming intended edits in final products that are released commercially. The limited amount of head-to-head comparisons of genome editing tools in diverse crops and livestock make it difficult to develop broad conclusions and best practices, which is further compounded by the diversity of techniques, targets, and processes. Developers and breeders should consult the literature and test as needed to determine which editing technology will be the most effective for their purposes, especially as more tools with altered efficiency and specificity become available. Yet, the lack of off-target edits in studies that employed careful design of targeting components followed by wide testing for on- and off-target edits bodes well for the use of genome editing with proper precautions of target selection and screening.

scholarly journals Establishment of a Genome Editing Tool Using CRISPR-Cas9 in Chlorella vulgaris UTEX395

2021 ◽  
Vol 22 (2) ◽  
pp. 480
Author(s):  
Jongrae Kim ◽  
Kwang Suk Chang ◽  
Sangmuk Lee ◽  
EonSeon Jin
Keyword(s):  
Genome Editing ◽  
Chlorella Vulgaris ◽  
Negative Selection ◽  
Screening Strategy ◽  
Feed Additive ◽  
Cell Factory ◽  
Dna Transformation ◽  
Research Groups ◽  
A Genome ◽  
Food And Feed

To date, Chlorella vulgaris is the most used species of microalgae in the food and feed additive industries, and also considered as a feasible cell factory for bioproducts. However, the lack of an efficient genetic engineering tool makes it difficult to improve the physiological characteristics of this species. Therefore, the development of new strategic approaches such as genome editing is trying to overcome this hurdle in many research groups. In this study, the possibility of editing the genome of C. vulgaris UTEX395 using clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) has been proven to target nitrate reductase (NR) and adenine phosphoribosyltransferase (APT). Genome-edited mutants, nr and apt, were generated by a DNA-mediated and/or ribonucleoprotein (RNP)-mediated CRISPR-Cas9 system, and isolated based on the negative selection against potassium chlorate or 2-fluoroadenine in place of antibiotics. The null mutation of edited genes was demonstrated by the expression level of the correspondent proteins or the mutation of transcripts, and through growth analysis under specific nutrient conditions. In conclusion, this study offers relevant empirical evidence of the possibility of genome editing in C. vulgaris UTEX395 by CRISPR-Cas9 and the practical methods. Additionally, among the generated mutants, nr can provide an easier screening strategy during DNA transformation than the use of antibiotics owing to their auxotrophic characteristics. These results will be a cornerstone for further advancement of the genetics of C. vulgaris.

Teaching Animals and Religion

2021 ◽  
Vol 25 (1) ◽  
pp. 48-70
Author(s):  
Dave Aftandilian
Keyword(s):  
Best Practices ◽  
Experiential Education ◽  
Final Section ◽  
Religious Studies ◽  
Contemplative Practices ◽  
Pros And Cons

Abstract Although animals have served as subjects and objects of religion since the Paleolithic, they are often omitted from standard religious studies courses. In this article, I discuss some best practices for introducing students to the study of animals and religion. After outlining some of the benefits of teaching about animals and religion, I explain the pros and cons of the two main approaches: by tradition or by topic. The majority of the article discusses some of the most important topics to include, as well as how best to approach several of them in terms of pedagogy and media. The final section explains the importance of bringing real animals into courses like this, and offers a variety of experiential education techniques for doing so, including contemplative practices.

A genome-editing off switch

2016 ◽  
Vol 18 (2) ◽  
pp. 69-69
Author(s):  
Ross Cloney
Keyword(s):  
Genome Editing ◽  
A Genome

Nanopore sequencing reveals a structural alteration of mirror‐image duplicated genes in a genome‐editing mouse line

2019 ◽  
Vol 60 (4) ◽  
pp. 120-125 ◽  
Author(s):  
Sachiko Miyamoto ◽  
Kazushi Aoto ◽  
Takuya Hiraide ◽  
Mitsuko Nakashima ◽  
Shuji Takabayashi ◽  
...  
Keyword(s):  
Genome Editing ◽  
Mirror Image ◽  
Structural Alteration ◽  
Nanopore Sequencing ◽  
Duplicated Genes ◽  
Mouse Line ◽  
A Genome

CRISPR/Cas9: a historical and chemical biology perspective of targeted genome engineering

2016 ◽  
Vol 45 (24) ◽  
pp. 6666-6684 ◽  
Author(s):  
Amrita Singh ◽  
Debojyoti Chakraborty ◽  
Souvik Maiti
Keyword(s):  
Genome Editing ◽  
Chemical Biology ◽  
Genome Engineering ◽  
A Genome

The development and adaptation of CRISPR–Cas9 as a genome editing tool and chemical biology approaches for modulating its activity.

scholarly journals A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy

2020 ◽  
Author(s):  
Jichao Wang ◽  
Xinyue Sui ◽  
Yamei Ding ◽  
Yingxin Fu ◽  
Xinjun Feng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Genome Editing ◽  
The Other ◽  
Plasmid Curing ◽  
Gene Deletions ◽  
A Genome ◽  
Single Clone ◽  
Previous Round ◽  
Genome Modifications

Abstract The production of optimized strains of a specific phenotype requires the construction and testing of a large number of genome modifications and combinations thereof. Most bacterial iterative genome-editing methods include essential steps to eliminate selection markers, or to cure plasmids. Additionally, the presence of escapers leads to time-consuming separate single clone picking and subsequent cultivation steps. Herein, we report a genome-editing method based on a Rock-Paper-Scissors (RPS) strategy. Each of three constructed sgRNA plasmids can cure, or be cured by, the other two plasmids in the system; plasmids from a previous round of editing can be cured while the current round of editing takes place. Due to the enhanced curing efficiency and embedded double check mechanism, separate steps for plasmid curing or confirmation are not necessary, and only two times of cultivation are needed per genome-editing round. This method was successfully demonstrated in Escherichia coli and Klebsiella pneumoniae with both gene deletions and replacements. To the best of our knowledge, this is the fastest and most robust iterative genome-editing method, with the least times of cultivation decreasing the possibilities of spontaneous genome mutations.

Sign in / Sign up

Export Citation Format

Share Document