scholarly journals CRISPR-Cas9 and beyond: what’s next in plant genome engineering

2021 ◽  
Author(s):  
Erin Zess ◽  
Matthew Begemann
Keyword(s):  
Real World ◽  
Genome Engineering ◽  
Agronomic Traits ◽  
Plant Genome ◽  
Zinc Finger Nucleases ◽  
Plant Genomes ◽  
Tal Effector ◽  
Real World Applications ◽  
Tal Effector Nucleases ◽  
Successive Generations

AbstractScientists have developed and deployed successive generations of genome engineering technologies for use in plants, including meganucleases, zinc finger nucleases, TAL effector nucleases, and CRISPR nucleases. Each of these tools has been hailed as potentially revolutionary, capable of providing more efficient and precise ways to modify plant genomes toward improving agronomic traits or making fundamental discoveries. The CRISPR nucleases, in particular, have accelerated the pace of innovation and expanded the boundaries of what is achievable within the plant research space. This review will take care to discuss current plant genome engineering technologies, covering both well-established and up-and-coming tools, as well as describe potential and real-world applications.

Genome Engineering with TAL-Effector Nucleases and Alternative Modular Nuclease Technologies

2013 ◽  
Vol 13 (4) ◽  
pp. 291-303 ◽  
Author(s):  
Andrew Scharenberg ◽  
Philippe Duchateau ◽  
Julianne Smith
Keyword(s):  
Genome Engineering ◽  
Tal Effector ◽  
Tal Effector Nucleases

scholarly journals Modern Trends in Plant Genome Editing: An Inclusive Review of the CRISPR/Cas9 Toolbox

2019 ◽  
Vol 20 (16) ◽  
pp. 4045 ◽  
Author(s):  
Ali Razzaq ◽  
Fozia Saleem ◽  
Mehak Kanwal ◽  
Ghulam Mustafa ◽  
Sumaira Yousaf ◽  
...  
Keyword(s):  
Genome Editing ◽  
Genome Engineering ◽  
Crop Improvement ◽  
Crop Plants ◽  
Targeted Mutagenesis ◽  
Plant Genome ◽  
Zinc Finger Nucleases ◽  
Base Substitution ◽  
Transcription Activator ◽  
Abiotic Stressors

Increasing agricultural productivity via modern breeding strategies is of prime interest to attain global food security. An array of biotic and abiotic stressors affect productivity as well as the quality of crop plants, and it is a primary need to develop crops with improved adaptability, high productivity, and resilience against these biotic/abiotic stressors. Conventional approaches to genetic engineering involve tedious procedures. State-of-the-art OMICS approaches reinforced with next-generation sequencing and the latest developments in genome editing tools have paved the way for targeted mutagenesis, opening new horizons for precise genome engineering. Various genome editing tools such as transcription activator-like effector nucleases (TALENs), zinc-finger nucleases (ZFNs), and meganucleases (MNs) have enabled plant scientists to manipulate desired genes in crop plants. However, these approaches are expensive and laborious involving complex procedures for successful editing. Conversely, CRISPR/Cas9 is an entrancing, easy-to-design, cost-effective, and versatile tool for precise and efficient plant genome editing. In recent years, the CRISPR/Cas9 system has emerged as a powerful tool for targeted mutagenesis, including single base substitution, multiplex gene editing, gene knockouts, and regulation of gene transcription in plants. Thus, CRISPR/Cas9-based genome editing has demonstrated great potential for crop improvement but regulation of genome-edited crops is still in its infancy. Here, we extensively reviewed the availability of CRISPR/Cas9 genome editing tools for plant biotechnologists to target desired genes and its vast applications in crop breeding research.

scholarly journals Genome Editing in Medicine: Tools and Challenges

2021 ◽  
Vol 28 (2) ◽  
pp. 8
Author(s):  
Gunda Petraitytė ◽  
Eglė Preikšaitienė ◽  
Violeta Mikštienė
Keyword(s):  
Genome Editing ◽  
Viral Vectors ◽  
Biological Therapy ◽  
Zinc Finger Nucleases ◽  
Target Cells ◽  
Biological Processes ◽  
Tal Effector ◽  
Bioethical Issues ◽  
Tal Effector Nucleases ◽  
Editing Process

Studies which seek fundamental, thorough knowledge of biological processes, and continuous advancement in natural sciences and biotechnology enable the establishment of molecular strategies and tools to treat disorders caused by genetic mutations. Over the years biological therapy evolved from using stem cells and viral vectors to RNA therapy and testing different genome editing tools as promising gene therapy agents. These genome editing technologies (Zinc finger nucleases, TAL effector nucleases), specifically CRISPR-Cas system, revolutionized the field of genetic engineering and is widely applied to create cell and animal models for various hereditary, infectious human diseases and cancer, to analyze and understand the molecular and cellular base of pathogenesis, to find potential drug/treatment targets, to eliminate pathogenic DNA changes in various medical conditions and to create future “precise medication”. Although different concerning factors, such as precise system delivery to the target cells, efficacy and accuracy of editing process, different approaches of making the DNA changes as well as worrying bioethical issues remain, the importance of genome editing technologies in medicine is undeniable. The future of innovative genome editing approach and strategies to treat diseases is complicated but interesting and exciting at once for all related parties – researchers, clinicians, and patients.

scholarly journals Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Yi Zhang ◽  
Zhen Liang ◽  
Yuan Zong ◽  
Yanpeng Wang ◽  
Jinxing Liu ◽  
...  
Keyword(s):  
Durum Wheat ◽  
Genome Editing ◽  
Bread Wheat ◽  
Transient Expression ◽  
Plant Species ◽  
Genome Engineering ◽  
Plant Genome ◽  
Engineering Research ◽  
Plant Genomes ◽  
Highly Efficient

Abstract Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.

222 PRODUCTION OF KNOCKOUT RATS BY USING ZINC FINGER NUCLEASES AND TAL EFFECTOR NUCLEASES

2014 ◽  
Vol 26 (1) ◽  
pp. 225
Author(s):  
T. Kaneko ◽  
T. Mashimo
Keyword(s):  
Embryo Transfer ◽  
Zinc Finger ◽  
Gene Locus ◽  
Genetically Engineered ◽  
Zinc Finger Nucleases ◽  
Rat Strains ◽  
Rat Embryos ◽  
Tal Effector ◽  
Tal Effector Nucleases

The rat has been used as an important animal for understanding human diseases. Genetically engineered rat strains are used as a human disease model in various research fields. Genetically engineered rat strains are now being routinely produced, not only as transgenic animals but also using gene knockout techniques. Recently, zinc finger nucleases (ZFN) and TAL effector nucleases (TALEN) have enabled editing targeted genes without using embryonic stem cells. These techniques have been applied for production of the knockout and knockin animals. We here studied that the effects of gene targeting by ZFN and TALEN introduced into rat embryos for efficient production of knockout rats. We custom-designed ZFN and TALEN plasmids targeted rat interleukin 2 receptor gamma (Il2rg) gene. Each mRNA was transcribed in vitro from these plasmids. Final concentration of mRNA was adjusted at 10 ng μL–1 in sterilized water for microinjection. Messenger RNA was injected into rat pronuclear stage embryos. The embryos were then cultured in vitro to the 2-cell stage, and were transferred into oviducts of pseudopregnant females. The rate of development of offspring of embryos and effects of editing targeted genes were examined. Of 41 two-cell embryos introduced ZFN after embryo transfer, 9 embryos (22%) developed to offspring. Three offspring (33%) had an edited targeted gene locus. In the embryos introduced TALEN, 30% (6 offspring) of embryos developed to offspring after embryo transfer and all offspring had an edited targeted gene locus. This study demonstrated that the ZFN and TALEN mRNA was active after introduction into rat embryos. Knockout rats could be produced by introduction of ZFN and TALEN into rat embryos. ZFN and TALEN will provide a powerful new approach for targeted gene editing not only in rats but also in other animal species.

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