scholarly journals Gene targeting technologies in rats: Zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats

2013 ◽  
Vol 56 (1) ◽  
pp. 46-52 ◽  
Author(s):  
Tomoji Mashimo
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases ◽  
Transcription Activator

scholarly journals Comparing Zinc Finger Nucleases and Transcription Activator-Like Effector Nucleases for Gene Targeting in Drosophila

2013 ◽  
Vol 3 (10) ◽  
pp. 1717-1725 ◽  
Author(s):  
Kelly J. Beumer ◽  
Jonathan K. Trautman ◽  
Michelle Christian ◽  
Timothy J. Dahlem ◽  
Cathleen M. Lake ◽  
...  
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases ◽  
Transcription Activator

scholarly journals Mini-Review Regarding the Applicability of Genome Editing Techniques Developed for Studying Infertility

Diagnostics ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 246
Author(s):  
Bogdan Doroftei ◽  
Ovidiu-Dumitru Ilie ◽  
Maria Puiu ◽  
Alin Ciobica ◽  
Ciprian Ilea
Keyword(s):  
Motor Activity ◽  
Experimental Model ◽  
Zinc Finger ◽  
Biomedical Research ◽  
Zinc Finger Nucleases ◽  
Loss Of Function ◽  
Transcription Activator ◽  
Phenotypic Changes ◽  
Wide Range ◽  
Short Palindromic Repeat

Infertility is a highly debated topic today. It has been long hypothesized that infertility has an idiopathic cause, but recent studies demonstrated the existence of a genetic substrate. Fortunately, the methods of editing the human genome proven to be revolutionary. Following research conducted, we identified a total of 21 relevant studies; 14 were performed on mice, 5 on zebrafish and 2 on rats. We concluded that over forty-four genes in total are dispensable for fertility in both sexes without affecting host homeostasis. However, there are genes whose loss-of-function induces moderate to severe phenotypic changes in both sexes. There were situations in which the authors reported infertility, exhibited by the experimental model, or other pathologies such as cryptorchidism, cataracts, or reduced motor activity. Overall, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 are techniques that offer a wide range of possibilities for studying infertility, even to create mutant variants. It can be concluded that ZFNs, TALENs, and CRISPR/Cas9 are crucial tools in biomedical research.

scholarly journals Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases

2008 ◽  
Vol 105 (50) ◽  
pp. 19821-19826 ◽  
Author(s):  
K. J. Beumer ◽  
J. K. Trautman ◽  
A. Bozas ◽  
J.-L. Liu ◽  
J. Rutter ◽  
...  
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases ◽  
Efficient Gene

Zinc finger nucleases-mediated gene targeting in plant cells

2010 ◽  
Vol 150 ◽  
pp. 474-475
Author(s):  
A Tovkach ◽  
V Zeevi ◽  
D Weinthal ◽  
T Tzfira
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Plant Cells ◽  
Zinc Finger Nucleases

Gene targeting using zinc finger nucleases

2005 ◽  
Vol 23 (8) ◽  
pp. 967-973 ◽  
Author(s):  
Matthew H Porteus ◽  
Dana Carroll
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases

Gene Targeting in Drosophila and Caenorhabditis elegans With Zinc-Finger Nucleases

2008 ◽  
pp. 63-77 ◽  
Author(s):  
Dana Carroll ◽  
Kelly J. Beumer ◽  
J. Jason Morton ◽  
Ana Bozas ◽  
Jonathan K. Trautman
Keyword(s):  
Caenorhabditis Elegans ◽  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases

236 GENE TARGETING USING ZINC FINGER NUCLEASES (ZFN) IN THE PORCINE FIBROBLAST

2012 ◽  
Vol 24 (1) ◽  
pp. 230
Author(s):  
S. Kim ◽  
J. W. Kim ◽  
S. M. Lee ◽  
J. H. Kim ◽  
M. J. Kang
Keyword(s):  
Homologous Recombination ◽  
Gene Targeting ◽  
Zinc Finger ◽  
Marker Gene ◽  
Somatic Cells ◽  
Embryonic Stem ◽  
Domestic Animals ◽  
Zinc Finger Nucleases ◽  
Exogenous Dna ◽  
Targeting Vector

Gene targeting is a genetic technique that utilises homologous recombination between an engineered exogenous DNA fragment and the endogenous genome of an animal. In domestic animals, gene targeting has provided an important tool for producing knockout pigs for the α1,3-galactosyltransferase gene (GGTA1) to use in xenotransplantation. The frequency of homologous recombination is a critical parameter for the success of gene targeting. The efficiency of homologous recombination in somatic cells is lower than that in mouse embryonic stem cells. The application of gene targeting to somatic cells has been limited by its low efficiency. Recently, knockout rat and mouse were generated by introducing nonhomologus end joining (NHE)-mediated deletion or insertion at the target site using zinc-finger nucleases (ZFN). Therefore, the development of effective knockout and knock-in techniques in domestic animals is very important in biomedical research. In this study, we investigated homologous recombination events at the cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene locus using ZFN in porcine primary fibroblast. The CMAH-targeted ZFN plasmid and mRNA were purchased from Sigma-Aldrich (St Louis, MO, USA). Porcine ear fibroblasts cells were obtained from a 10-day-old male Chicago miniature pigs. The fibroblasts were cultured in DMEM containing 15% fetal bovine serum, 1 × nonessential amino acids, 1 × sodium pyruvate, 10–4 M β-mercaptoethanol, 100 unit mL–1 penicillin and 100 μg mL–1 streptomycin. The cells were trypsinized and resuspended at a concentration of 1.25 × 107 cells mL–1 in F10 nutrient mixture. Four hundred microliters of the cell suspension was electroporated in a 4-mm cuvette with 4 pulses of 1 ms duration using 400V capacitive discharges using the CMAH neo targeting vector and ZFN plasmid or RNA. The CMAH neo targeting vector consists of the neomycin resistance gene (neo) as a positive selectable marker gene, 789-bp 5′ arm and 763-bp 3′ arm from exon 8 of CMAH gene. After selection of G-418, PCR analysis was performed using 64 colonies transfected with ZFN plasmid and 48 colonies transfected with ZFN RNA. As a result, 19 positive colonies were identified in colonies transfected with ZFN plasmid and 15 colonies were identified in colonies transfected with ZFN RNA. The targeting efficiency was 29.7 and 31.6% in the colonies transfected with ZFN plasmid and ZFN RNA, respectively. To our knowledge, this study provides the first evidence that the efficiency of gene targeting using ZFN was higher than that of conventional gene targeting in the porcine fibroblast. These cell lines may be used in production of CMAH knockouts for xenotransplantation.

scholarly journals Gene targeting to the ROSA26 locus directed by engineered zinc finger nucleases

2011 ◽  
Vol 40 (8) ◽  
pp. 3741-3752 ◽  
Author(s):  
Pablo Perez-Pinera ◽  
David G. Ousterout ◽  
Matthew T. Brown ◽  
Charles A. Gersbach
Keyword(s):  
Gene Targeting ◽  
Zinc Finger ◽  
Zinc Finger Nucleases ◽  
Rosa26 Locus

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.

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