CRISPR/Cas system research has advanced significantly in biological sciences.There are still many challenges to effective delivery before efficient gene editing may be achieved

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Plant Breeding ◽  
Rapid Growth ◽  
Gene Editing ◽  
Life Sciences ◽  
Great Promise ◽  
System Research ◽  
Efficient Gene ◽  
Effective Delivery ◽  
Agricultural Applications ◽  
Substantial Progress

The CRISPR/Cas system's discovery and execution offer great promise for the treatment of human illnesses and the revolutionization of plant breeding. Despite the fact that CRISPR/Cas system research is well advanced in the life sciences community, there are still considerable barriers to effective delivery that must be overcome before effective gene editing can be achieved. Consider characteristics like specificity, efficacy, and controlled expression while deciding on a plan. Due to the discovery of innovative delivery systems, many of the shortcomings of the CRISPR/Cas system's translatability have been addressed. Because of the rapid growth of delivery mechanisms, a successful translation of CRISPR/Cas technology into medical and agricultural applications is vital, and substantial progress is predicted.

scholarly journals Progress in Gene-Editing Technology of Zebrafish

Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1300
Author(s):  
Yanling Li ◽  
Zhipeng Jia ◽  
Shuchao Zhang ◽  
Xiaozhen He
Keyword(s):  
Danio Rerio ◽  
Gene Editing ◽  
Life Sciences ◽  
Strand Break ◽  
Vital Role ◽  
Genome Modification ◽  
Efficient Gene ◽  
Vertebrate Model ◽  
Multi Method Approach ◽  
Method Approach

As a vertebrate model, zebrafish (Danio rerio) plays a vital role in the field of life sciences. Recently, gene-editing technology has become increasingly innovative, significantly promoting scientific research on zebrafish. However, the implementation of these methods in a reasonable and accurate manner to achieve efficient gene-editing remains challenging. In this review, we systematically summarize the development and latest progress in zebrafish gene-editing technology. Specifically, we outline trends in double-strand break-free genome modification and the prospective applications of fixed-point orientation transformation of any base at any location through a multi-method approach.

Genome Editing with CRISPR

2016 ◽  
Vol 41 (3) ◽  
pp. 1-3
Author(s):  
Nicanor Pier Giorgio Austriaco ◽  
Keyword(s):  
Genetic Engineering ◽  
Genome Editing ◽  
Gene Editing ◽  
Life Sciences ◽  
Great Promise ◽  
Bacterial Cells ◽  
Kitchen Table ◽  
A Genome ◽  
Do It Yourself

There has been much discussion regarding the proper use of the powerful CRISPR technologies that can be used to edit the genome. CRISPR is a technique borrowed from bacterial cells that will allow scientists to quickly and precisely change the DNA of nearly any organism, including humans. Unlike other gene-editing technologies, CRISPR is cheap, quick, and easy to use. In fact, do-it-yourself CRISPR genome editing kits are available online for less than $200, which will enable anyone, including so-called biohackers, to do genetic engineering at the kitchen table. In only three years—CRISPR as a genome-editing tool was first described in 2012—it is already universally acknowledged that this technology will revolutionize the life sciences. But CRISPR’s great promise has also sparked a great ethical and societal debate on its legitimate uses, most significantly on whether it should be used to alter the genomes of our children and grandchildren.

scholarly journals Simplified All-In-One CRISPR-Cas9 Construction for Efficient Genome Editing in Cryptococcus Species

2021 ◽  
Vol 7 (7) ◽  
pp. 505
Author(s):  
Ping Zhang ◽  
Yu Wang ◽  
Chenxi Li ◽  
Xiaoyu Ma ◽  
Lan Ma ◽  
...  
Keyword(s):  
Genome Editing ◽  
Fungal Pathogens ◽  
Gene Editing ◽  
Recombination Frequency ◽  
Target Sequence ◽  
Widespread Application ◽  
Genetic Studies ◽  
Efficient Gene ◽  
Cryptococcus Species ◽  
Overlap Pcr

Cryptococcus neoformans and Cryptococcus deneoformans are opportunistic fungal pathogens found worldwide that are utilized to reveal mechanisms of fungal pathogenesis. However, their low homologous recombination frequency has greatly encumbered genetic studies. In preliminary work, we described a ‘suicide’ CRISPR-Cas9 system for use in the efficient gene editing of C. deneoformans, but this has not yet been used in the C. neoformans strain. The procedures involved in constructing vectors are time-consuming, whether they involve restriction enzyme-based cloning of donor DNA or the introduction of a target sequence into the gRNA expression cassette via overlap PCR, as are sophisticated, thus impeding their widespread application. Here, we report the optimized and simplified construction method for all-in-one CRISPR-Cas9 vectors that can be used in C. neoformans and C. deneoformans strains respectively, named pNK003 (Genbank: MW938321) and pRH003 (Genbank: KX977486). Taking several gene manipulations as examples, we also demonstrate the accuracy and efficiency of the new simplified all-in-one CRISPR-Cas9 genome editing tools in both Serotype A and Serotype D strains, as well as their ability to eliminate Cas9 and gDNA cassettes after gene editing. We anticipate that the availability of new vectors that can simplify and streamline the technical steps for all-in-one CRISPR-Cas9 construction could accelerate genetic studies of the Cryptococcus species.

scholarly journals Human Genetic Diversity Alters Therapeutic Gene Editing Off-Target Outcomes

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3993-3993
Author(s):  
Linda Yingqi Lin ◽  
Samuele Cancellieri ◽  
Jing Zeng ◽  
Francesco Masillo ◽  
My Anh Nguyen ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Clinical Trials ◽  
Genome Editing ◽  
Gene Editing ◽  
Conflicts Of Interest ◽  
African Ancestry ◽  
Great Promise ◽  
Personal Genome ◽  
Hematopoietic Stem ◽  
The Impact

Abstract CRISPR gene editing holds great promise to modify somatic genomes to ameliorate disease. In silico prediction of homologous sites coupled with biochemical evaluation of possible genomic off-targets may predict genotoxicity risk of individual gene editing reagents. However, standard computational and biochemical methods focus on reference genomes and do not consider the impact of genetic diversity on off-target potential. Here we developed a web application called CRISPRme that explicitly and efficiently integrates human genetic variant datasets with orthogonal genomic annotations to predict and prioritize off-target sites at scale. The method considers both single-nucleotide variants (SNVs) and indels, accounts for bona fide haplotypes, accepts spacer:protospacer mismatches and bulges, and is suitable for personal genome analyses. We tested the tool with a guide RNA (gRNA) targeting the BCL11A erythroid enhancer that has shown therapeutic promise in clinical trials for sickle cell disease (SCD) and β-thalassemia (Frangoul et al. NEJM 2021). We find that the top predicted off-target site is produced by a non-reference allele common in African-ancestry populations (rs114518452, minor allele frequency (MAF) = 4.5%) that introduces a protospacer adjacent motif (PAM) for SpCas9. We validate that SpCas9 generates indels (~9.6% frequency) and chr2 pericentric inversions in a strictly allele-specific manner in edited CD34+ hematopoietic stem/progenitor cells (HSPCs), although a high-fidelity Cas9 variant mitigates this off-target. This report illustrates how population and private genetic variants should be considered as modifiers of genome editing outcomes. We expect that variant-aware off-target assessment will be required for therapeutic genome editing efforts going forward, including both ongoing and future clinical trials, and we provide a powerful approach for comprehensive off-target prediction. CRISPRme is available at crisprme.di.univr.it. Disclosures No relevant conflicts of interest to declare.

scholarly journals Design of time-delayed safety switches for CRISPR gene therapy

2021 ◽  
Author(s):  
Dashan Sun
Keyword(s):  
Gene Therapy ◽  
Immune Response ◽  
Time Delay ◽  
Cell Lines ◽  
Gene Editing ◽  
Drug Accumulation ◽  
Work Time ◽  
Crispr System ◽  
Efficient Gene ◽  
Target Effect

CRISPR system is a powerful gene editing tool which has already been reported to address a variety of gene relevant diseases in different cell lines. However, off-target effect and immune response caused by Cas9 remain two fundamental problems. In our work, time-delayed safety switches are designed based on either artificial ultrasensitivity transmission module or intrinsic time delay in biomolecular activities. By addressing gene therapy efficiency, off-target effect, immune response and drug accumulation, we hope our safety switches may offer inspiration in realizing safe and efficient gene therapy in humans.

scholarly journals All Plant Breeding Technologies Are Equal, but Some Are More Equal Than Others: The Case of GM and Mutagenesis

2021 ◽  
Vol 12 ◽  
Author(s):  
Luisa Batalha ◽  
Francesco Foroni ◽  
Brian Joseph Jones
Keyword(s):  
Risk Perception ◽  
Plant Breeding ◽  
Genetic Modification ◽  
Public Perception ◽  
Gene Editing ◽  
Consumer Attitudes ◽  
Food Items ◽  
Gm Foods ◽  
Genetic Development ◽  
Sexual Crosses

A pervasive opposition to genetically modified (GM) foods has developed from the notion that they pose a risk to human and environmental health. Other techniques for the genetic modification of plants, such as sexual crossing and mutagenesis breeding, have mostly remained unchallenged. This research aims to investigate public perception of plant breeding technologies. Specifically, sexual crossing, mutagenesis, transgenics (GM) and gene editing. It was expected that attitudes and intentions would be most positive and the perception of risk lowest for plant genetic modification through sexual crosses. Scores on these variables were expected to be similar between mutagenesis, GM and gene editing. It was also expected that attitudes, intentions and risk perception would change (becoming more positive) once participants learned about foods developed through these technologies. Participants reported their attitudes, intentions and risk perception at two points in time. At Time 2, they were presented with pictures of food items developed through sexual crossing, GM and mutagenesis. The results showed that mutagenesis stood out as the most negatively perceived technology, whereas genetic development via sexual crosses was generally perceived as positive. The results highlight the importance of messaging, framing in consumer attitudes.

Plant breeding for resilient pastures

2021 ◽  
Vol 17 ◽  
Author(s):  
John Caradus ◽  
Joseph Bouton ◽  
Charles Brummer ◽  
Marty Faville ◽  
Richard George ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Plant Breeding ◽  
Seed Yield ◽  
Abiotic Stresses ◽  
Dry Matter ◽  
Gene Editing ◽  
Nutritive Value ◽  
Environmental Concerns ◽  
Biotic And Abiotic Stresses ◽  
Plant Introductions

Plant breeding has had, and continues to have, an important role in providing farmers with resilient pastures. Early breeding relied on improvement of ecotype populations and this was accelerated by crossing with selected introduced germplasm. The primary traits under selection have targeted speed of establishment, total and/or seasonal dry matter (DM) yield, nutritive value or feed quality, flowering time and reduced aftermath heading, disease resistance, persistence and seed yield. Continued improvement through plant breeding to meet environmental concerns and tolerances to both biotic and abiotic stresses will be achieved through ongoing plant introductions, exploiting heterosis, speed breeding, genomic selection, improvements in phenotyping, metabolomics, improved compatibility with beneficial microbes, and potentially the use of transgenic and gene editing technologies.

Cas9 conjugate complex delivering donor DNA for efficient gene editing by homology-directed repair

2021 ◽  
Author(s):  
Yoo Kyung Kang ◽  
Ju Hee Lee ◽  
San Hae Im ◽  
Joo Hoon Lee ◽  
Juhee Jeong ◽  
...  
Keyword(s):  
Gene Editing ◽  
Homology Directed Repair ◽  
Efficient Gene

scholarly journals Recent Approaches to Chiral 1,4-Dihydropyridines and their Fused Analogues

Catalysts ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1019
Author(s):  
Martins Rucins ◽  
Aiva Plotniece ◽  
Eiva Bernotiene ◽  
Wei-Bor Tsai ◽  
Arkadij Sobolev
Keyword(s):  
Kinetic Resolution ◽  
Stereoselective Synthesis ◽  
Biological Activities ◽  
Great Promise ◽  
Racemic Mixtures ◽  
Treatment Of Hypertension ◽  
Opposite Action ◽  
Recent Developments ◽  
Substantial Progress ◽  
Active Calcium

The purpose of this review is to highlight recent developments in the synthesis of chiral 1,4-dihydropyridines and their fused analogues. 1,4-Dihydropyridines are among the most active calcium antagonists that are used for the treatment of hypertension. Enantiomers of unsymmetrical 1,4-dihydropyridines often show different biological activities and may have even an opposite action profile. Hantzsch synthesis usually produces racemic mixtures of unsymmetrical 1,4-dihydropyridines. Therefore, the development of stereoselective synthesis of 1,4-dihydropyridines is one of the priorities of medicinal chemistry. Over the years, numerous methodologies have been developed for the production of enantiopure 1,4-dihydropyridines, such as stereoselective synthesis using chiral auxiliaries and chiral cyclocondensation partners, chromatographical methods, resolution of diastereomeric 1,4-dihydropyridine salts, enzyme catalysed kinetic resolution, or asymmetrisation of ester groups of 1,4-dihydropyridines. These approaches have been studied in detail and are relatively well established. The catalytic asymmetric approach holds the greatest promise in delivering the most practical and widely applicable methods. Substantial progress has been made toward the development of enantioselective organocatalytic methods for the construction of the chiral dihydropyridines. However, most of them do not provide a convenient way to pharmacologically important 1,4-dihydropyridine-3,5-dicarboxylates. Organocatalytic enantioselective desymmetrisation of prochiral 1,4-dihydropyridine-3,5-dicarbaldehydes also has great promise in the synthesis of pharmacologically important 1,4-dihydropyridine-3,5-dicarboxylates.

scholarly journals Gene Therapy: The Molecular Bandage for Treating Genetic Disorders

1970 ◽  
Vol 3 (1) ◽  
pp. 24-27
Author(s):  
Md Manjurul Karim
Keyword(s):  
Gene Therapy ◽  
Viral Vectors ◽  
Genetic Disorders ◽  
Clinical Status ◽  
Genetic Material ◽  
Review Article ◽  
Cell Tissue ◽  
Efficient Gene ◽  
Effective Delivery ◽  
A Cell

The concept of gene therapy involves the transfer of genetic material into a cell, tissue, or whole organ, with a view to curing a disease or at least improving the clinical status of a patient. Much of its success relies heavily on the development of an effective delivery system that is capable of efficient gene transfer in a variety of tissues, without causing any associated pathogenic effects. Viral vectors currently offer the best choice for efficient gene delivery, what has been discussed in this review article. Their performance and pathogenecity has been evaluated in animal models, and encouraging results form the basis for clinical trials to treat genetic disorders and acquired diseases. Despite some initial success in these trials, vector development remains a seminal concern for improved gene therapy technologies. DOI: http://dx.doi.org/10.3329/akmmcj.v3i1.10110 AKMMCJ 2012; 3(1): 24-27

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