Identification of Off-Target Cleavage Sites of Zinc Finger Nucleases and TAL Effector Nucleases Using Predictive Models

2014 ◽  
pp. 371-383 ◽  
Author(s):  
Eli J. Fine ◽  
Thomas J. Cradick ◽  
Gang Bao
Keyword(s):  
Zinc Finger ◽  
Predictive Models ◽  
Zinc Finger Nucleases ◽  
Cleavage Sites ◽  
Tal Effector ◽  
Tal Effector Nucleases

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.

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 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.

scholarly journals Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases

2012 ◽  
Vol 3 (1) ◽  
Author(s):  
Takahito Watanabe ◽  
Hiroshi Ochiai ◽  
Tetsushi Sakuma ◽  
Hadley W. Horch ◽  
Naoya Hamaguchi ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Tal Effector ◽  
Hemimetabolous Insect ◽  
Tal Effector Nucleases ◽  
Genome Modifications
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