scholarly journals Can pseudocomplementary peptide nucleic acid nucleases (pcPNANs) be a new tool for genetic engineering?

2014 ◽  
Author(s):  
Penghui Shi
Keyword(s):  
Nucleic Acid ◽  
Genetic Engineering ◽  
Peptide Nucleic Acid ◽  
Cleavage Site ◽  
New Technologies ◽  
Selection Procedure ◽  
Zinc Finger Nucleases ◽  
Biological Research ◽  
Target Sequence ◽  
Transcription Activator

Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) comprise a powerful class of tools that are redefining the boundaries of biological research. Although these technologies have begun to enable targeted genome modifications, there remains a need for new technologies that are scalable, affordable, and easy to engineer. In this paper, we propose a new tool for genetic engineering, the pseudocomplementary peptide nucleic acid nucleases (pcPNANs), which are composed of a pseudocomplementary PNA (pcPNA) specific for a DNA target sequence, a FokI nuclease cleavage domain and a nuclear localization signal. pcPNANs may induce targeted DNA double-strand breaks that activate DNA damage response pathways and enable custom alterations. Their cleavage-site is determined by simple Watson-Crick rule, and thus pcPNANs for aimed cleavage of genomes can be straightforwardly designed and synthesized without any selection procedure. Accordingly, the cleavage-site and site-specificity are freely chosen by changing the sequences and the lengths of pcPNA strands. We believe that the potentiality of pcPNAN as a new tool for genetic engineering will be confirmed in the future.

scholarly journals Can pseudocomplementary peptide nucleic acid nucleases (pcPNANs) be a new tool for genetic engineering?

2014 ◽  
Author(s):  
Penghui Shi
Keyword(s):  
Nucleic Acid ◽  
Genetic Engineering ◽  
Peptide Nucleic Acid ◽  
Cleavage Site ◽  
New Technologies ◽  
Selection Procedure ◽  
Zinc Finger Nucleases ◽  
Biological Research ◽  
Target Sequence ◽  
Transcription Activator

Zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) comprise a powerful class of tools that are redefining the boundaries of biological research. Although these technologies have begun to enable targeted genome modifications, there remains a need for new technologies that are scalable, affordable, and easy to engineer. In this paper, we propose a new tool for genetic engineering, the pseudocomplementary peptide nucleic acid nucleases (pcPNANs), which are composed of a pseudocomplementary PNA (pcPNA) specific for a DNA target sequence, a FokI nuclease cleavage domain and a nuclear localization signal. pcPNANs may induce targeted DNA double-strand breaks that activate DNA damage response pathways and enable custom alterations. Their cleavage-site is determined by simple Watson-Crick rule, and thus pcPNANs for aimed cleavage of genomes can be straightforwardly designed and synthesized without any selection procedure. Accordingly, the cleavage-site and site-specificity are freely chosen by changing the sequences and the lengths of pcPNA strands. We believe that the potentiality of pcPNAN as a new tool for genetic engineering will be confirmed in the future.

scholarly journals Exploiting DNA Endonucleases to Advance Mechanisms of DNA Repair

Biology ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 530
Author(s):  
Marlo K. Thompson ◽  
Robert W. Sobol ◽  
Aishwarya Prakash
Keyword(s):  
Dna Repair ◽  
Mammalian Cells ◽  
Genome Stability ◽  
Gene Editing ◽  
Adaptive Immune System ◽  
Zinc Finger Nucleases ◽  
Specific Gene ◽  
Target Sequence ◽  
Transcription Activator ◽  
Low Cytotoxicity

The earliest methods of genome editing, such as zinc-finger nucleases (ZFN) and transcription activator-like effector nucleases (TALENs), utilize customizable DNA-binding motifs to target the genome at specific loci. While these approaches provided sequence-specific gene-editing capacity, the laborious process of designing and synthesizing recombinant nucleases to recognize a specific target sequence, combined with limited target choices and poor editing efficiency, ultimately minimized the broad utility of these systems. The discovery of clustered regularly interspaced short palindromic repeat sequences (CRISPR) in Escherichia coli dates to 1987, yet it was another 20 years before CRISPR and the CRISPR-associated (Cas) proteins were identified as part of the microbial adaptive immune system, by targeting phage DNA, to fight bacteriophage reinfection. By 2013, CRISPR/Cas9 systems had been engineered to allow gene editing in mammalian cells. The ease of design, low cytotoxicity, and increased efficiency have made CRISPR/Cas9 and its related systems the designer nucleases of choice for many. In this review, we discuss the various CRISPR systems and their broad utility in genome manipulation. We will explore how CRISPR-controlled modifications have advanced our understanding of the mechanisms of genome stability, using the modulation of DNA repair genes as examples.

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.

Modern biotechnology tools and biosecurity in the Middle East

2021 ◽  
Vol 16 (11) ◽  
pp. 155-163
Author(s):  
Alsubki Roua
Keyword(s):  
Middle East ◽  
Genetic Engineering ◽  
Genome Editing ◽  
Middle Eastern ◽  
Strand Break ◽  
Biological Weapons ◽  
Zinc Finger Nucleases ◽  
Transcription Activator ◽  
Human Genomes ◽  
Modern Biotechnology

The global health system is under a constant threat from microbial outbreaks. The innovation in genetic engineering has created an existential threat to national, regional and international security. This threat, that can edit microbial or human genomes, requires global attention. In the current review, a comprehensive literature search was conducted using PubMed, SCOPUS and Google Scholar to identify literature discussing modern biotechnology tools as well as relevance to biosafety in the Middle east region. This review was undertaken to provide an overview of biological threats due to advancements in genetic engineering, making it possible to insert or delete specific genes to increase the virulence of particular microbes. These pathogens or other toxic factors can be multiplied by technology, creating new biological weapons. Genome editing technologies including meganucleases (MNs), zinc finger nucleases (ZFNs), transcription activator-like effector (TALE)-nucleases (TALENs) and recently discovered clustered regularly interspaced short palindromic repeats (CRISPR/Cas) induce a double strand break at specific DNA target site. Genome editing technologies lead to an irreversible and permanent alteration of the genetic code and therefore, can inevitably result in security risks. Vulnerabilities in Middle Eastern laboratories raise the prospect of high levels of pathogenic microbes potentially creating a weakness in the diagnosis and monitoring of epidemics. Furthermore, the lack of regional legislation to regulate biosafety and biosecurity may lead to biological threat at the regional level.

scholarly journals Application of genome-editing systems to enhance available pig resources for agriculture and biomedicine

2020 ◽  
Vol 32 (2) ◽  
pp. 40
Author(s):  
Kiho Lee ◽  
Kayla Farrell ◽  
Kyungjun Uh
Keyword(s):  
Genetic Engineering ◽  
Genome Editing ◽  
Direct Injection ◽  
Somatic Cells ◽  
Cost Effective ◽  
Genetically Engineered ◽  
Zinc Finger Nucleases ◽  
Transcription Activator ◽  
Developmental Defects ◽  
Site Specific

Traditionally, genetic engineering in the pig was a challenging task. Genetic engineering of somatic cells followed by somatic cell nuclear transfer (SCNT) could produce genetically engineered (GE) pigs carrying site-specific modifications. However, due to difficulties in engineering the genome of somatic cells and developmental defects associated with SCNT, a limited number of GE pig models were reported. Recent developments in genome-editing tools, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system, have markedly changed the effort and time required to produce GE pig models. The frequency of genetic engineering in somatic cells is now practical. In addition, SCNT is no longer essential in producing GE pigs carrying site-specific modifications, because direct injection of genome-editing systems into developing embryos introduces targeted modifications. To date, the CRISPR/Cas9 system is the most convenient, cost-effective, timely and commonly used genome-editing technology. Several applicable biomedical and agricultural pig models have been generated using the CRISPR/Cas9 system. Although the efficiency of genetic engineering has been markedly enhanced with the use of genome-editing systems, improvements are still needed to optimally use the emerging technology. Current and future advances in genome-editing strategies will have a monumental effect on pig models used in agriculture and biomedicine.

scholarly journals Principles of Genetic Engineering

Genes ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 291 ◽  
Author(s):  
Thomas M. Lanigan ◽  
Huira C. Kopera ◽  
Thomas L. Saunders
Keyword(s):  
Homologous Recombination ◽  
Genetic Engineering ◽  
Dna Sequences ◽  
Cultured Cells ◽  
Embryonic Stem ◽  
Zinc Finger Nucleases ◽  
Chromosome Break ◽  
Transcription Activator ◽  
Random Integration ◽  
Dna Insertion

Genetic engineering is the use of molecular biology technology to modify DNA sequence(s) in genomes, using a variety of approaches. For example, homologous recombination can be used to target specific sequences in mouse embryonic stem (ES) cell genomes or other cultured cells, but it is cumbersome, poorly efficient, and relies on drug positive/negative selection in cell culture for success. Other routinely applied methods include random integration of DNA after direct transfection (microinjection), transposon-mediated DNA insertion, or DNA insertion mediated by viral vectors for the production of transgenic mice and rats. Random integration of DNA occurs more frequently than homologous recombination, but has numerous drawbacks, despite its efficiency. The most elegant and effective method is technology based on guided endonucleases, because these can target specific DNA sequences. Since the advent of clustered regularly interspaced short palindromic repeats or CRISPR/Cas9 technology, endonuclease-mediated gene targeting has become the most widely applied method to engineer genomes, supplanting the use of zinc finger nucleases, transcription activator-like effector nucleases, and meganucleases. Future improvements in CRISPR/Cas9 gene editing may be achieved by increasing the efficiency of homology-directed repair. Here, we describe principles of genetic engineering and detail: (1) how common elements of current technologies include the need for a chromosome break to occur, (2) the use of specific and sensitive genotyping assays to detect altered genomes, and (3) delivery modalities that impact characterization of gene modifications. In summary, while some principles of genetic engineering remain steadfast, others change as technologies are ever-evolving and continue to revolutionize research in many fields.

Role of stem cells in large animal genetic engineering in the TALENs–CRISPR era

2014 ◽  
Vol 26 (1) ◽  
pp. 65 ◽  
Author(s):  
Ki-Eun Park ◽  
Bhanu Prakash V. L. Telugu
Keyword(s):  
Stem Cells ◽  
Genetic Engineering ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Zinc Finger Nucleases ◽  
Large Animal ◽  
Prime Model ◽  
Transcription Activator

The establishment of embryonic stem cells (ESCs) and gene targeting technologies in mice has revolutionised the field of genetics. The relative ease with which genes can be knocked out, and exogenous sequences introduced, has allowed the mouse to become the prime model for deciphering the genetic code. Not surprisingly, the lack of authentic ESCs has hampered the livestock genetics field and has forced animal scientists into adapting alternative technologies for genetic engineering. The recent discovery of the creation of induced pluripotent stem cells (iPSCs) by upregulation of a handful of reprogramming genes has offered renewed enthusiasm to animal geneticists. However, much like ESCs, establishing authentic iPSCs from the domestic animals is still beset with problems, including (but not limited to) the persistent expression of reprogramming genes and the lack of proven potential for differentiation into target cell types both in vitro and in vivo. Site-specific nucleases comprised of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulated interspaced short palindromic repeats (CRISPRs) emerged as powerful genetic tools for precisely editing the genome, usurping the need for ESC-based genetic modifications even in the mouse. In this article, in the aftermath of these powerful genome editing technologies, the role of pluripotent stem cells in livestock genetics is discussed.

scholarly journals CRISPR-Cas Genome Editing Tool: Mechanisms of Pathogen Resistance Plants – Review

2019 ◽  
Vol 7 ◽  
pp. 69-80
Author(s):  
Karthik Kalidoss
Keyword(s):  
Genome Editing ◽  
Plant Protection ◽  
Site Directed Mutagenesis ◽  
Blast Disease ◽  
Experimental Methodology ◽  
Zinc Finger Nucleases ◽  
Biological Research ◽  
Transcription Activator ◽  
Fungus Infection ◽  
Exposure Site

In recent years, the CRISPR-Cas system is most familiar and advance genome editing tool in modern biological research. The genome editing tool used in various biological researchers worldwide in past years has witnessed exposure site-directed mutagenesis modification methods zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), Meganucleases and CRISPR-Cas9(associated proteins 9). CRISPR-Cas genome editing technology to ease design and implement, more flexible and less expensive. Plants are affected two types of stresses like biotic and abiotic. Abiotic occurs naturally temperature or wind, sunlight depend upon on the environmental conditions. Biotic stress is caused by pathogens of virus, fungi, bacteria, etc. This review to focus on the recent advance of plant protection use CRISPR-Cas system mechanism of disease resistant plants in past and current trends of research. A short overview of the experimental methodology for Beet Curly Top Virus (BCTV) disease and Magnaporthe oryzae fungus infection cause rice blast disease resistance mechanisms will be discussed. Furthermore, the need developments of this genome editing tool in future.

scholarly journals Genome Editing: A New Horizon for Oral and Craniofacial Research

2018 ◽  
Vol 98 (1) ◽  
pp. 36-45 ◽  
Author(s):  
N. Yu ◽  
J. Yang ◽  
Y. Mishina ◽  
W.V. Giannobile
Keyword(s):  
Genome Editing ◽  
Genetic Manipulation ◽  
Zinc Finger Nucleases ◽  
Biological Research ◽  
Transcription Activator ◽  
Developmental Defects ◽  
Genetic Manipulations ◽  
Research Fields ◽  
Repair Mechanisms ◽  
Conducting Research

Precise and efficient genetic manipulations have enabled researchers to understand gene functions in disease and development, providing a platform to search for molecular cures. Over the past decade, the unprecedented advancement of genome editing techniques has revolutionized the biological research fields. Early genome editing strategies involved many naturally occurring nucleases, including meganucleases, zinc finger nucleases, and transcription activator-like effector-based nucleases. More recently, the clustered regularly interspaced short palindromic repeats (CRISPR) / CRISPR-associated nucleases (CRISPR/Cas) system has greatly enriched genetic manipulation methods in conducting research. Those nucleases generate double-strand breaks in the target gene sequences and then utilize DNA repair mechanisms to permit precise yet versatile genetic manipulations. The oral and craniofacial field harbors a plethora of diseases and developmental defects that require genetic models that can exploit these genome editing techniques. This review provides an overview of the genome editing techniques, particularly the CRISPR/Cas9 technique, for the oral and craniofacial research community. We also discuss the details about the emerging applications of genome editing in oral and craniofacial biology.

Molecular insights on the dynamic stability of peptide nucleic acid functionalized carbon and boron nitride nanotubes

2021 ◽  
Vol 23 (1) ◽  
pp. 219-228
Author(s):  
Nabanita Saikia ◽  
Mohamed Taha ◽  
Ravindra Pandey
Keyword(s):  
Nucleic Acid ◽  
Boron Nitride ◽  
Nucleic Acids ◽  
Dynamic Stability ◽  
Peptide Nucleic Acid ◽  
Rational Design ◽  
Peptide Nucleic Acids ◽  
Boron Nitride Nanotubes ◽  
Molecular Hybrids ◽  
Single Wall Nanotubes

The rational design of self-assembled nanobio-molecular hybrids of peptide nucleic acids with single-wall nanotubes rely on understanding how biomolecules recognize and mediate intermolecular interactions with the nanomaterial's surface.

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