scholarly journals Practical Approaches for Knock-Out Gene Editing in Pigs

2021 ◽  
Vol 11 ◽  
Author(s):  
Laura Daniela Ratner ◽  
Gaston Emilio La Motta ◽  
Olinda Briski ◽  
Daniel Felipe Salamone ◽  
Rafael Fernandez-Martin
Keyword(s):  
Nuclear Transfer ◽  
Gene Editing ◽  
Early Stage ◽  
Human Diseases ◽  
Meat Production ◽  
Porcine Genome ◽  
Knock Out ◽  
The Past ◽  
Programmable Nucleases ◽  
First Time

Pigs are an important resource for meat production and serve as a model for human diseases. Due to their physiological and anatomical similarities to humans, these animals can recapitulate symptoms of human diseases, becoming an effective model for biomedical research. Although, in the past pig have not been widely used partially because of the difficulty in genetic modification; nowadays, with the new revolutionary technology of programmable nucleases, and fundamentally of the CRISPR-Cas9 systems, it is possible for the first time to precisely modify the porcine genome as never before. To this purpose, it is necessary to introduce the system into early stage zygotes or to edit cells followed by somatic cell nuclear transfer. In this review, several strategies for pig knock-out gene editing, using the CRISPR-Cas9 system, will be summarized, as well as genotyping methods and different delivery techniques to introduce these tools into the embryos. Finally, the best approaches to produce homogeneous, biallelic edited animals will be discussed.

scholarly journals Animal cloning by nuclear transfer: state-of-the-art and future perspectives.

2005 ◽  
Vol 52 (3) ◽  
pp. 585-588 ◽  
Author(s):  
Andras Dinnyes ◽  
Agnes Szmolenszky
Keyword(s):  
Nuclear Transfer ◽  
Genetic Basis ◽  
State Of The Art ◽  
New Technology ◽  
Transgenic Animal ◽  
Human Diseases ◽  
Genomic Research ◽  
Model Organisms ◽  
Future Perspectives ◽  
Knock Out

Model organisms are essential to study the genetic basis of human diseases. Transgenic mammalian models, especially genetic knock-out mice have catalysed the progress in this area. To continue the advancement, further sophisticated and refined models are crucially needed to study the genetic basis and manifestations of numerous human diseases. Coinciding with the start of the new era of post-genomic research, new tools for establishment of transgenesis, such as nuclear transfer and gene targeting in somatic cells, have become available, offering a unique opportunity for the generation of transgenic animal models. The new technology provides important tools for comparative functional genomics to promote the interpretation and increase the practical value of the data generated in numerous mouse models. This paper discusses the state-of-the-art of the nuclear replacement technology and presents future perspectives.

scholarly journals CRISPR as a Potential Technique to Reduce Suicide Risk

2021 ◽  
Vol 10 (3) ◽  
Author(s):  
Hidemi Zamora ◽  
Javier Cornejo
Keyword(s):  
Cystic Fibrosis ◽  
Suicide Risk ◽  
Gene Editing ◽  
Design Tool ◽  
High Importance ◽  
Knock Out ◽  
Guide Rna ◽  
Guide Rnas ◽  
The Past ◽  
Rna Design

As suicide is the nineteenth leading cause of death worldwide, it is important to focus on discovering ways to reduce the risk of suicide-related death as much as possible. With CRISPR starting to become increasingly popular over the past few years, this gene editing technique has been used to study how to edit, turn off, or knock out multiple parts of the genome. However, research on genes related to diseases as cystic fibrosis or Alzheimer’s disease has been mainly prioritized and, even though they are of high importance as well, important issues such as suicide have been left into oblivion. Four genes have been proven to be key in influencing suicide risk, showing that not only environmental factors account for an increased possibility of death by this cause. Therefore, gene editing techniques such as CRISPR could be applied in order to knock out those genes and reduce this risk. This research used Synthego’s guide RNA design tool to predict how the use of CRISPR can be helpful in knocking out those four suicide-related genes and, consequently, in preventing suicide. The top-ranked guide RNAs for each gene were used, showing the best results possible and with the least number of off-targets, which, in turn, demonstrates the effectiveness of CRISPR as a potential technique to reduce the number of suicide-related deaths worldwide.

scholarly journals Effective MSTN Gene Knockout by AdV-Delivered CRISPR/Cas9 in Postnatal Chick Leg Muscle

2020 ◽  
Vol 21 (7) ◽  
pp. 2584 ◽  
Author(s):  
Ke Xu ◽  
Cheng Xiao Han ◽  
Hao Zhou ◽  
Jin Mei Ding ◽  
Zhong Xu ◽  
...  
Keyword(s):  
Growth And Development ◽  
Gene Editing ◽  
Gene Knockout ◽  
Muscle Growth ◽  
Meat Production ◽  
Wild Type ◽  
Knock Out ◽  
Mstn Gene ◽  
Differentiation And Proliferation ◽  
Transcriptomic Changes

Muscle growth and development are important aspects of chicken meat production, but the underlying regulatory mechanisms remain unclear and need further exploration. CRISPR has been used for gene editing to study gene function in mice, but less has been done in chick muscles. To verify whether postnatal gene editing could be achieved in chick muscles and determine the transcriptomic changes, we knocked out Myostatin (MSTN), a potential inhibitor of muscle growth and development, in chicks and performed transcriptome analysis on knock-out (KO) muscles and wild-type (WT) muscles at two post-natal days: 3d (3-day-old) and 14d (14-day-old). Large fragment deletions of MSTN (>5 kb) were achieved in all KO muscles, and the MSTN gene expression was significantly downregulated at 14d. The transcriptomic results indicated the presence of 1339 differentially expressed genes (DEGs) between the 3d KO and 3d WT muscles, as well as 597 DEGs between 14d KO and 14d WT muscles. Many DEGs were found to be related to cell differentiation and proliferation, muscle growth and energy metabolism. This method provides a potential means of postnatal gene editing in chicks, and the results presented here could provide a basis for further investigation of the mechanisms involved in muscle growth and development.

scholarly journals Generation of biallelic knock-out sheep via gene-editing and somatic cell nuclear transfer

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Honghui Li ◽  
Gui Wang ◽  
Zhiqiang Hao ◽  
Guozhong Zhang ◽  
Yubo Qing ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Gene Editing ◽  
Knock Out

scholarly journals Production of a Cloned Offspring and CRISPR/Cas9 Genome Editing of Embryonic Fibroblasts in Cattle

2021 ◽  
Vol 496 (1) ◽  
pp. 48-51
Author(s):  
G. N. Singina ◽  
P. V. Sergiev ◽  
A. V. Lopukhov ◽  
M. P. Rubtsova ◽  
N. P. Taradajnic ◽  
...  
Keyword(s):  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Gene Editing ◽  
Protein Gene ◽  
Fibroblast Cell ◽  
Fibroblast Cells ◽  
Fibroblast Cell Line ◽  
Embryonic Fibroblasts ◽  
Knock Out ◽  
Beta Lactoglobulin

Abstract Somatic Cell Nuclear Transfer (SCNT) technique was used to produce the first viable cloned cattle offspring in Russia. Whole-genome SNP genotyping confirmed that the cloned calf was identical to the fibroblast cell line that was used for SCNT. CRISPR/Cas9 approach was subsequently used to knock out genes for beta-lactoglobulin gene (PAEP) and the beta-lactoglobulin-like protein gene (LOC100848610) in the fibroblast cells. Gene editing (GE) efficiency was 4.4% for each of these genes. We successfully obtained single-cell-derived fibroblast colonies containing PAEP and LOC100848610 knockouts, which will be used to produce beta-lactoglobulin-deficient cattle.

‘Higher then to her no bookes doe reach’

2018 ◽  
Author(s):  
Martha Vandrei
Keyword(s):  
Early Stage ◽  
Deep Understanding ◽  
Contemporary Philosophy ◽  
James I ◽  
Historical Method ◽  
The Past ◽  
Classical World ◽  
Material Evidence ◽  
Do So

This chapter and the following both draw the reader into seventeenth-century understandings of the past, and of Boudica in particular, and makes clear that in a time before disciplines, writers of ‘history’ were erudite commentators, immersed in political thought, the classical world, and contemporary ideas, as well as in drama, poetry, and the law. Chapter 1 shows the subtleties of Boudica’s place in history at this early stage by giving sustained attention to the work of Edmund Bolton (1574/5–c.1634), the first person to analyse the written and material evidence for Boudica’s deeds, and the last to do so in depth before the later nineteenth century. Bolton’s distaste for contemporary philosophy and his loyalty to James I were highly influential in determining the way the antiquary approached Boudica and her rebellion; but equally important was Bolton’s deep understanding of historical method and the strictures this placed on his interpretive latitude.

scholarly journals Efficient TALEN-mediated gene knockin at the bovine Y chromosome and generation of a sex-reversal bovine

2021 ◽  
Author(s):  
Ming Wang ◽  
ZhaoLin Sun ◽  
Fangrong Ding ◽  
Haiping Wang ◽  
Ling Li ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Nuclear Transfer ◽  
Sex Reversal ◽  
Practical Application ◽  
Transcription Activator ◽  
New Methods ◽  
Fetal Fibroblasts ◽  
Male Phenotype ◽  
Solid Foundation ◽  
First Time

AbstractFunctional elucidation of bovine Y-chromosome genes requires available genome editing technologies. Meanwhile, it has yet to be proven whether the bovine Sry gene is the main or single factor involved in the development of the male phenotype in bovine. Here, we efficiently knocked out four Y-linked genes (Sry, ZFY, DDX3Y, and EIF2S3Y) in bovine fetal fibroblasts (BFFs) with transcription activator-like effector nucleases (TALENs) individually. Furthermore, we used TALEN-mediated gene knockin at the Sry gene and generated a sex-reversal bovine by somatic cell nuclear transfer (SCNT). The resulting bovine had only one ovary and was sterile. We demonstrate, for the first time, that the Sry gene is an important sex-determining gene in bovine. Our method lays a solid foundation for detecting the biology of the bovine Y chromosome, as it may provide an alternative biological model system for the study of mammalian sex determination, and new methods for the practical application in agricultural, especially for sex predetermination.

scholarly journals DNA–protein crosslink proteases in genome stability

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Annamaria Ruggiano ◽  
Kristijan Ramadan
Keyword(s):  
Dna Replication ◽  
Cancer Therapy ◽  
Genome Stability ◽  
Dna Lesions ◽  
Protein Component ◽  
Human Diseases ◽  
Direct Link ◽  
The Past ◽  
Protein Crosslinks ◽  
Bulky Dna Lesions

AbstractProteins covalently attached to DNA, also known as DNA–protein crosslinks (DPCs), are common and bulky DNA lesions that interfere with DNA replication, repair, transcription and recombination. Research in the past several years indicates that cells possess dedicated enzymes, known as DPC proteases, which digest the protein component of a DPC. Interestingly, DPC proteases also play a role in proteolysis beside DPC repair, such as in degrading excess histones during DNA replication or controlling DNA replication checkpoints. Here, we discuss the importance of DPC proteases in DNA replication, genome stability and their direct link to human diseases and cancer therapy.

FC14-01 - Internal losses and the creation of “Nowhere land”: psychodynamic themes of adults with ADHD discovered during drama-therapy

2011 ◽  
Vol 26 (S2) ◽  
pp. 1893-1893
Author(s):  
I. Manor ◽  
G. Yazpan
Keyword(s):  
Drama Therapy ◽  
Transitional Space ◽  
Men And Women ◽  
The Past ◽  
Chronic Disorder ◽  
Pharmacological Aspect ◽  
Inner Self ◽  
Socioeconomic Strata ◽  
The Creation ◽  
First Time

ADHD is a well-known, chronic disorder that persists in adulthood. During the past 20 years its existence in adults is becoming clearer, yet its dynamic aspects are rarely discussed. The treatment of adults is vital, as much as that of children; however the literature discussing it, especially its non-pharmacological aspect, is scarce.We describe the results of our treatment with drama-therapy of two groups of adults with ADHD. These groups included 11 adults (from both groups), men and women, from most socioeconomic strata, aged ≥ 60 yrs., who were diagnosed as suffering from ADHD and were treated for it for the first time in their life. Drama-therapy was selected as we believed it to be a useful method with associative, distracted ADHD patients, since it enabled the use of transitional space through non-verbal images and acts.This presentation discusses the basic themes with which patients began therapy. Interestingly, all patients, however different, shared the same themes that were built on self doubt and the pre-presumption of disappointment. The impairment related to ADHD, that was felt, but not understood, led to a strong experience of heavy losses, which we tried to define separately: of a clear path, of control, of the inner perception of borders and of the loss of an integrative inner self. All these losses were accumulated in the transitional space in a place we named “Nowhere land”.We would like to present these themes of losses and of becoming lost and to discuss their meaning.

BRAZILIAN BISHOP OUTLINES "A PASTORAL STRATEGY IN TIMES OF REVOLUTION"

Exchange ◽  
1982 ◽  
Vol 11 (1) ◽  
pp. 48-57
Keyword(s):  
Northeast Brazil ◽  
Profound Change ◽  
Christian Communities ◽  
The Past ◽  
First Time

AbstractMons. Antonio Batista Fragoso has been bishop of Crateus in Northeast Brazil for the past ten years. Eighty percent of the 360,000 people in his diocese are impoverished peasants who engage in rudimentary farming. At least half of the peasants are landless. It is among these people that Bishop Fragoso has encouraged the formation of small grassroots Christian communities that are responsible for a profound change in the patterns of Christian living in his diocese. The following is LP's translation of excerpts from a talk that Bishop Fragoso gave to his fellow bishops and priests in Managua in October, 1980. This is the first time it has appeared in English. (Editor Latinamerica Press)

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