Molecular Aspects of Zinc Finger Nucleases (ZFNs)-Mediated Gene Editing in Rat Embryos

2018 ◽  
pp. 295-312
Author(s):  
Xiaoxia Cui
Keyword(s):  
Zinc Finger ◽  
Gene Editing ◽  
Zinc Finger Nucleases ◽  
Rat Embryos ◽  
Molecular Aspects

scholarly journals Gene Editing of Human Embryonic Stem Cells via an Engineered Baculoviral Vector Carrying Zinc-finger Nucleases

2011 ◽  
Vol 19 (5) ◽  
pp. 942-950 ◽  
Author(s):  
Yuning Lei ◽  
Chi-Lin Lee ◽  
Kye-Il Joo ◽  
Jonathan Zarzar ◽  
Yarong Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Human Embryonic Stem Cells ◽  
Gene Editing ◽  
Embryonic Stem ◽  
Zinc Finger Nucleases

scholarly journals THE JOURNEY OF CRISPR-CAS9 FROM BACTERIAL DEFENSE MECHANISM TO A GENE EDITING TOOL IN BOTH ANIMALS AND PLANTS

2020 ◽  
Vol 2020 (1) ◽  
Author(s):  
T Tahir ◽  
Q Ali ◽  
MS Rashid ◽  
A Malik
Keyword(s):  
Immune System ◽  
Genetic Engineering ◽  
Success Rate ◽  
Genome Editing ◽  
Zinc Finger ◽  
Gene Editing ◽  
Defense Mechanism ◽  
Zinc Finger Nucleases ◽  
Transcription Activator

Today we can use multiple of endonucleases for genome editing which has become very important and used in number of applications. We use sequence specific molecular scissors out of which, most important are mega nucleases, zinc finger nucleases, TALENS (Transcription Activator Like-Effector Nucleases) and CRISPR-Cas9 which is currently the most famous due to a number of reasons, they are cheap, easy to build, very specific in nature and their success rate in plants and animals is also high. Who knew that one day these CRISPR discovered as a part of immune system of bacteria will be this much worthwhile in the field of genetic engineering? This review interprets the science behind their mechanism and how several advancements were made with the passage of time to make them more efficient for the assigned job.

scholarly journals Delivery Approaches for Therapeutic Genome Editing and Challenges

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1113 ◽  
Author(s):  
Ilayda Ates ◽  
Tanner Rathbone ◽  
Callie Stuart ◽  
P. Hudson Bridges ◽  
Renee N. Cottle
Keyword(s):  
Clinical Trials ◽  
Genome Editing ◽  
Zinc Finger ◽  
Gene Editing ◽  
Zinc Finger Nucleases ◽  
Systemic Delivery ◽  
Transcription Activator ◽  
Therapeutic Gene ◽  
Major Barrier ◽  
Associated Proteins

Impressive therapeutic advances have been possible through the advent of zinc-finger nucleases and transcription activator-like effector nucleases. However, discovery of the more efficient and highly tailorable clustered regularly interspaced short palindromic repeats (CRISPR) and associated proteins (Cas9) has provided unprecedented gene-editing capabilities for treatment of various inherited and acquired diseases. Despite recent clinical trials, a major barrier for therapeutic gene editing is the absence of safe and effective methods for local and systemic delivery of gene-editing reagents. In this review, we elaborate on the challenges and provide practical considerations for improving gene editing. Specifically, we highlight issues associated with delivery of gene-editing tools into clinically relevant cells.

Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery

2008 ◽  
Vol 40 (2) ◽  
pp. 278 ◽  
Author(s):  
A. Lombardo ◽  
P. Genovese ◽  
C.M. Beausejour ◽  
S. Colleoni ◽  
Y.-L. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Zinc Finger ◽  
Gene Editing ◽  
Lentiviral Vector ◽  
Zinc Finger Nucleases ◽  
Human Stem Cells ◽  
Vector Delivery

Editing genetico: nuova questione bioetica? / Gene editing: a new issue for Bioethics?

2017 ◽  
Vol 66 (3) ◽  
pp. 291-304 ◽  
Author(s):  
Pietro Refolo ◽  
Vincenzo L. Pascali ◽  
Antonio G. Spagnolo
Keyword(s):  
Present Article ◽  
Zinc Finger ◽  
Gene Editing ◽  
Point Of View ◽  
Zinc Finger Nucleases ◽  
Ethical Point

Modificazioni controllate nel genoma sono possibili, tramite svariate tecniche, sin dagli anni ’70. Nucleasi a dito di zinco, nucleasi TALE, ma soprattutto CRISPR-Cas9 sono tecniche di editing genetico che hanno reso più semplice effettuarle. Il sistema CRISPR-Cas9, in particolare, si sta dimostrando estremamente vantaggioso in termini di accessibilità, efficienza e versatilità. Gli obiettivi del presente contributo consistono nel: 1. ricostruire i “fatti” salienti che hanno determinato l’emergere del topic dell’“editing genetico”; 2. provare a dar risposta a un primo fondamentale interrogativo circa l’originalità dei dilemmi etici da esso sollevato con particolare riferimento al CRISPR-Cas9. La conclusione è che, allo stato attuale, l’impiego di queste nuove tecniche non solleva questioni etiche nuove. L’unica eccezione sembrerebbe essere data dal particolare tipo di mutazioni indotte da queste tecniche, indistinguibili a lungo termine da quelle prodotte dalla natura, fatto che sta già determinando qualche difficoltà nella classificazione degli OGM ottenuti tramite queste tecniche. ---------- Controlled genome mutations are made possible through several techniques since the ’70s. Zinc finger nucleases, TALE nucleases and above all CRISPR-Cas9 system are “gene editing” techniques which have made mutations easier. Particularly, CRISPR-Cas9 system seems to be extremely profitable in terms of accessibility, efficiency and versatility. The aims of the present article are: 1. to reconstruct the main “facts” about the birth of the topic on “gene editing”; to seek to answer a first question about the novelty of issues raised by this topic. Our conclusion is that, from an ethical point of view, using these techniques does not raise new ethical questions. Perhaps, the only exception refers to the specific mutations produced through these techniques which cannot be distinguished from natural mutations and makes GMO classification more difficult.

Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery

2007 ◽  
Vol 25 (11) ◽  
pp. 1298-1306 ◽  
Author(s):  
Angelo Lombardo ◽  
Pietro Genovese ◽  
Christian M Beausejour ◽  
Silvia Colleoni ◽  
Ya-Li Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Zinc Finger ◽  
Gene Editing ◽  
Lentiviral Vector ◽  
Zinc Finger Nucleases ◽  
Human Stem Cells ◽  
Vector Delivery

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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