scholarly journals Myoediting: Toward Prevention of Muscular Dystrophy by Therapeutic Genome Editing

2018 ◽  
Vol 98 (3) ◽  
pp. 1205-1240 ◽  
Author(s):  
Yu Zhang ◽  
Chengzu Long ◽  
Rhonda Bassel-Duby ◽  
Eric N. Olson
Keyword(s):  
Genome Editing ◽  
Genome Engineering ◽  
Genetic Disorders ◽  
Premature Death ◽  
Muscular Dystrophies ◽  
Genetic Mutations ◽  
Secondary Effects ◽  
Disease Treatment ◽  
Programmable Nucleases

Muscular dystrophies represent a large group of genetic disorders that significantly impair quality of life and often progress to premature death. There is no effective treatment for these debilitating diseases. Most therapies, developed to date, focus on alleviating the symptoms or targeting the secondary effects, while the underlying gene mutation is still present in the human genome. The discovery and application of programmable nucleases for site-specific DNA double-stranded breaks provides a powerful tool for precise genome engineering. In particular, the CRISPR/Cas system has revolutionized the genome editing field and is providing a new path for disease treatment by targeting the disease-causing genetic mutations. In this review, we provide a historical overview of genome-editing technologies, summarize the most recent advances, and discuss potential strategies and challenges for permanently correcting genetic mutations that cause muscular dystrophies.

Therapeutic genome editing with engineered nucleases

2017 ◽  
Vol 37 (01) ◽  
pp. 45-52 ◽  
Author(s):  
Simone Haas ◽  
Viviane Dettmer ◽  
Toni Cathomen
Keyword(s):  
Gene Therapy ◽  
Genome Editing ◽  
Genetic Disorders ◽  
Zinc Finger Nucleases ◽  
New Era ◽  
Engineered Nucleases ◽  
Genetic Interventions ◽  
Type Gene ◽  
Programmable Nucleases ◽  
Cancer Immunotherapies

SummaryTargeted genome editing with designer nucleases, such as zinc finger nucleases, TALE nucleases, and CRISPR-Cas nucleases, has heralded a new era in gene therapy. Genetic disorders, which have not been amenable to conventional gene-addition-type gene therapy approaches, such as disorders with dominant inheritance or diseases caused by mutations in tightly regulated genes, can now be treated by precise genome surgery. Moreover, engineered nucleases enable novel genetic interventions to fight infectious diseases or to improve cancer immunotherapies. Here, we review the development of the different classes of programmable nucleases, discuss the challenges and improvements in translating gene editing into clinical use, and give an outlook on what applications can expect to enter the clinic in the near future.

scholarly journals One-plasmid based CRISPR-Cas9 editing for gene deletion in Lactococcus lactis

2021 ◽  
Author(s):  
José Miguel Miquelão Santos ◽  
Gabriel António Amaro Monteiro ◽  
Duarte Miguel de França Teixeira dos Prazeres ◽  
Sofia de Oliveira Dias Duarte
Keyword(s):  
Genome Editing ◽  
Lactococcus Lactis ◽  
Plasmid Dna ◽  
Genome Engineering ◽  
Gene Deletion ◽  
Deletion Mutants ◽  
Cell Factories ◽  
Yield And Quality ◽  
Delivery Vehicles

Abstract Lactococcus lactis strains are promising cell factories and delivery vehicles of plasmid DNA and recombinant protein for therapeutic applications. However, the limited yields of recombinant molecules obtained with these bacteria limits their wide applicability. Genome engineering of this host may solve the problem. However, the current genome editing toolbox available for L. lactis is either too laborious or incapable of large edits, limiting the scope of strain editing experiments. In this work, the basis for a one-plasmid CRISPR-Cas9 based genome editing plasmid was developed and tested. The new plasmid (pTCas9dO) adapted from the pKCcas9dO plasmid was used to delete 657 bp of the lactococcal nuclease nth of L. lactis subsp. lactis LMG19460, with the aim of improving yield and quality of plasmid DNA replicated in this strain. Although deletion mutants were successfully generated, plasmid curing was unsuccessful. Thus, further modifications are required before the plasmid is truly applicable for genome editing experiments. Unexpectedly, the generated deletion mutants generated a roughly 40% decrease in plasmid yield alongside with a decrease in the quality of produced pDNA.

scholarly journals A genetically encoded inhibitor of 53BP1 to stimulate homology-based gene editing

2016 ◽  
Author(s):  
Marella D. Canny ◽  
Leo C.K. Wan ◽  
Amélie Fradet-Turcotte ◽  
Alexandre Orthwein ◽  
Nathalie Moatti ◽  
...  
Keyword(s):  
Genome Editing ◽  
Genome Engineering ◽  
Cell Types ◽  
Strand Break ◽  
End Joining ◽  
Dsb Repair ◽  
Non Homologous End Joining ◽  
G1 Cells ◽  
Programmable Nucleases ◽  
End Resection

AbstractThe expanding repertoire of programmable nucleases such as Cas9 brings new opportunities in genetic medicine1–3. In many cases, these nucleases are engineered to induce a DNA double-strand break (DSB) to stimulate precise genome editing by homologous recombination (HR). However, HR efficiency is nearly always hindered by competing DSB repair pathways such as non-homologous end-joining (NHEJ). HR is also profoundly suppressed in non-replicating cells, thus precluding the use of homology-based genome engineering in a wide variety4 of cell types. Here, we report the development of a genetically encoded inhibitor of 53BP1 (known as TP53BP1), a regulator of DSB repair pathway choice5. 53BP1 promotes NHEJ over HR by suppressing end resection, the formation of 3-prime single-stranded DNA tails, which is the rate-limiting step in HR initiation. 53BP1 also blocks the recruitment of the HR factor BRCA1 to DSB sites in G1 cells4,6. The inhibitor of 53BP1 (or i53) was identified through the screening of a massive combinatorial library of engineered ubiquitin variants by phage display7. i53 binds and occludes the ligand binding site of the 53BP1 Tudor domain with high affinity and selectivity, blocking its ability to accumulate at sites of DNA damage. i53 is a potent selective inhibitor of 53BP1 and enhances gene targeting and chromosomal gene conversion, two HR-dependent reactions. Finally, i53 can also activate HR in G1 cells when combined with the activation of end-resection and KEAP1 inhibition. We conclude that 53BP1 inhibition is a robust tool to enhance precise genome editing by canonical HR pathways.

Comparative Pharmacognostic, Physicochemical and Phytochemical Studies of Different Samples of Plumbago zeylanica L. Roots

2017 ◽  
Vol 9 (06) ◽  
Author(s):  
Neha Jain ◽  
Mohan Lal Kori
Keyword(s):  
Phytochemical Analysis ◽  
Local Market ◽  
Herbal Drugs ◽  
Disease Treatment ◽  
Organoleptic Evaluation ◽  
Plumbago Zeylanica ◽  
Qualitative And Quantitative ◽  
Crude Drugs ◽  
Phytochemical Studies

The objective of the present study is to evaluate the quality of the marketed and self collected samples of Plumbago zeylanica L. roots on the standardization parameters. This study is planned mainly to confirm changes with quality of drug. Now a day’s more demand of herbal drugs for disease treatment, lack of knowledge of proper methodology and availability are promoting the practices of adulteration and substitution. Thus, the standardization of the plant crude drugs is necessary to maintain their therapeutic efficacy. Comparative studies were carried out to evaluate the standards of P. zeylanica L. with emphasis on organoleptic evaluation, physicochemical and phytochemical analysis. Samples were procured from local market and self collected to determine the qualitative and quantitative variations. The result indicates that self collected sample showed significant results with comparison to marketed sample.

Evaluation of medical care for Diabetic and Hypertensive patients in primary care in Mexico: Observational retrospective study (Preprint)

2020 ◽  
Author(s):  
Agustin Lara-Esqueda ◽  
Sergio A Zaizar-Fregoso ◽  
Violeta M Madrigal-Perez ◽  
Mario Ramirez-Flores ◽  
Daniel A Montes-Galindo ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Medical Care ◽  
Serum Lipids ◽  
Quality Care ◽  
Premature Death ◽  
Current Standard ◽  
Anonymized Data ◽  
Quality Of Medical Care

BACKGROUND Diabetes Mellitus is a worldwide health problem and the leading cause of premature death with increasing prevalence over time. Usually, along with it, Hypertension presents and acts as another risk factor that increases mortality risk. Both diseases impact the country's health while also producing an economic burden for society, causing billions of dollars to be invested in their management. OBJECTIVE The present study evaluated the quality of medical care for patients diagnosed with diabetes mellitus (DM), hypertension (HBP), and both pathologies (DM+HBP) within a public health system in Mexico, according to the official Mexican standard for each pathology. METHODS 45,498 patients were included from 2012 to 2015. All information was taken from the electronic medical records database, exported as anonymized data for research purposes. Each patient record was compared against the standard to test the quality of medical care. RESULTS Glycemia with hypertension goals reached 29.6% in DM+HBP, 48.6% in DM, and 53.2% in HBP. The goals of serum lipids were reached by 3% in DM+HBP, 5% in DM, and 0.2% in HBP. Glycemia, hypertension, and LDL cholesterol reached 0.04%. 15% of patients had an undiagnosed disease of diabetes or hypertension. Clinical follow-up examinations reached 20% for foot examination and clinical eye examination in the whole population. Specialty referral reached 1% in angiology or cardiology in the whole population. CONCLUSIONS Goals for glycemic and hypertension reached 50% in the overall population, while serum lipids, clinical follow-up examinations, and referral to a specialist were deficient. Patients who had both diseases had more consultations, better control for hypertension and lipids, but inferior glycemic control. Overall, quality care for DM and/or HBP has not been met according to the standards. While patients with DM and HBP do not have a current standard to evaluate their own needs.

scholarly journals Annexins and Membrane Repair Dysfunctions in Muscular Dystrophies

2021 ◽  
Vol 22 (10) ◽  
pp. 5276
Author(s):  
Coralie Croissant ◽  
Romain Carmeille ◽  
Charlotte Brévart ◽  
Anthony Bouter
Keyword(s):  
Skeletal Muscle ◽  
Muscular Dystrophy ◽  
Genetic Disorders ◽  
Muscle Cells ◽  
Cell Types ◽  
Clinical Severity ◽  
Skeletal Muscle Cells ◽  
Muscular Dystrophies ◽  
Membrane Repair ◽  
Human Skeletal Muscle Cells

Muscular dystrophies constitute a group of genetic disorders that cause weakness and progressive loss of skeletal muscle mass. Among them, Miyoshi muscular dystrophy 1 (MMD1), limb girdle muscular dystrophy type R2 (LGMDR2/2B), and LGMDR12 (2L) are characterized by mutation in gene encoding key membrane-repair protein, which leads to severe dysfunctions in sarcolemma repair. Cell membrane disruption is a physiological event induced by mechanical stress, such as muscle contraction and stretching. Like many eukaryotic cells, muscle fibers possess a protein machinery ensuring fast resealing of damaged plasma membrane. Members of the annexins A (ANXA) family belong to this protein machinery. ANXA are small soluble proteins, twelve in number in humans, which share the property of binding to membranes exposing negatively-charged phospholipids in the presence of calcium (Ca2+). Many ANXA have been reported to participate in membrane repair of varied cell types and species, including human skeletal muscle cells in which they may play a collective role in protection and repair of the sarcolemma. Here, we discuss the participation of ANXA in membrane repair of healthy skeletal muscle cells and how dysregulation of ANXA expression may impact the clinical severity of muscular dystrophies.

scholarly journals Recent advances in CRISPR/Cas9 and applications for wheat functional genomics and breeding

aBIOTECH ◽  
2021 ◽  
Author(s):  
Jun Li ◽  
Yan Li ◽  
Ligeng Ma
Keyword(s):  
Functional Genomics ◽  
Genome Editing ◽  
Functional Annotation ◽  
Genome Engineering ◽  
Agronomic Traits ◽  
Wheat Breeding ◽  
Breeding Programs ◽  
Base Editing ◽  
The World ◽  
World Food Security

AbstractCommon wheat (Triticum aestivum L.) is one of the three major food crops in the world; thus, wheat breeding programs are important for world food security. Characterizing the genes that control important agronomic traits and finding new ways to alter them are necessary to improve wheat breeding. Functional genomics and breeding in polyploid wheat has been greatly accelerated by the advent of several powerful tools, especially CRISPR/Cas9 genome editing technology, which allows multiplex genome engineering. Here, we describe the development of CRISPR/Cas9, which has revolutionized the field of genome editing. In addition, we emphasize technological breakthroughs (e.g., base editing and prime editing) based on CRISPR/Cas9. We also summarize recent applications and advances in the functional annotation and breeding of wheat, and we introduce the production of CRISPR-edited DNA-free wheat. Combined with other achievements, CRISPR and CRISPR-based genome editing will speed progress in wheat biology and promote sustainable agriculture.

scholarly journals Fabry disease, a complex pathology not easy to diagnose

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Paolo Colomba ◽  
Simone Scalia ◽  
Giuseppe Cammarata ◽  
Carmela Zizzo ◽  
Daniele Francofonte ◽  
...  
Keyword(s):  
Fabry Disease ◽  
Vascular Endothelium ◽  
Clinical Manifestations ◽  
Premature Death ◽  
Lysosomal Storage ◽  
Familial Data ◽  
Enzyme Replacement ◽  
Cerebrovascular Complications ◽  
Cellular Events

Fabry disease is a multisystemic lysosomal storage disorder, inherited in an X-linked manner. It is a defect of metabolism of the glycosphingolipids, due to the reduction or absence of the activity of lysosomal enzyme α-galactosidase A. This reduction of activity causes the storage of globotriaosylceramide and derivatives in the lysosomes, triggering a cascade of cellular events, mainly in vascular endothelium. These events are the responsible for the systemic clinical manifestations and the renal, cardiac and cerebrovascular complications, or a combination of them. The symptomatology can lead to the premature death of patient between the fourth or fifth decade of life. The first symptoms can occur at different ages, generally in childhood, with different severity and course. Fabry disease is suspected on the basis of clinical and anamnestic-familial data, and it is confirmed by enzymatic and genetic assays. However, Fabry disease could be a pathology more complex than previously considered, and the diagnostic tests that are currently in use could be not always sufficient to confirm the clinical diagnosis. Probably, other factors could be also involved in the onset of symptomatology. In the last years, the knowledge of the disease is considerably increased but other studies are necessary to make a prompt and reliable diagnosis. An early diagnosis of Fabry disease is essential for the beginning of the enzyme replacement therapy, which can contribute to arrest its progression and improve the quality of life of patients.

scholarly journals Pattern of Hypertension in Fayoum Governorate

2019 ◽  
pp. 1-8
Author(s):  
Eman M. Ezzat ◽  
Maher A. Al Amir ◽  
Abdelrahman A. Ewais
Keyword(s):  
Prevalence Rate ◽  
Premature Death ◽  
Hypertensive Patients ◽  
Related Risk ◽  
High Prevalence ◽  
Study Population ◽  
Laboratory Investigations ◽  
Low Levels ◽  
Fayoum Governorate

Background: Hypertension is a preventable cause of premature death worldwide. Data from the Egyptian National Hypertension Project (NHP) revealed that hypertension is a significant problem among Egyptians. We studied the prevalence of hypertension, related risk factors and its complications in Fayoum governorate. Patients and Methods: This study included 2800 Egyptian participants aged 18 to 60 years old from Fayoum governorate selected randomly from different places in Fayoum governorate. Blood pressure was measured for them. Re-measurement for those with BP measurements ≥ 140/90 was done with a further assessment. Known hypertensive patients were thoroughly evaluated by clinical examination and laboratory investigations. Results: Our survey revealed that the prevalence rate of hypertension in Fayoum governorate was 28.4% (19.3% were aware and 9.1% were first diagnosed and unaware of their hypertension), 11% of the study population were pre-hypertensives. Poorly controlled hypertensive patients were 59.9%. Of all hypertensive patients, 63.4% of them were females, 87.9% of them were overweight and obese, while 26.3% of them had diabetes mellitus. Also, 41% of known hypertensive patients had complications. Conclusion and Recommendations: There is a high prevalence rate of hypertension in Fayoum governorate. More knowledge is required to reveal the reasons behind this high prevalence in addition to the low levels of control and awareness of hypertension, to put the appropriate strategy to improve the rate of control, awareness, and quality of life of hypertensive patients in Egypt.

scholarly journals CRISPR-Cas12a (Cpf1): A Versatile Tool in the Plant Genome Editing Tool Box for Agricultural Advancement

2020 ◽  
Author(s):  
Anindya Bandyopadhyay ◽  
Nagesh Kancharla ◽  
vivek javalkote ◽  
santanu dasgupta ◽  
Thomas Brutnell
Keyword(s):  
Genome Editing ◽  
Genome Engineering ◽  
Temperature Stability ◽  
Double Strand Breaks ◽  
Plant Genome ◽  
Strand Breaks ◽  
Guide Rna ◽  
Versatile Tool ◽  
Type V ◽  
Global Population

Global population is predicted to approach 10 billion by 2050, an increase of over 2 billion from today. To meet the demands of growing, geographically and socio-economically diversified nations, we need to diversity and expand agricultural production. This expansion of agricultural productivity will need to occur under increasing biotic, and environmental constraints driven by climate change. Clustered regularly interspaced short palindromic repeats-site directed nucleases (CRISPR-SDN) and similar genome editing technologies will likely be key enablers to meet future agricultural needs. While the application of CRISPR-Cas9 mediated genome editing has led the way, the use of CRISPR-Cas12a is also increasing significantly for genome engineering of plants. The popularity of the CRISPR-Cas12a, the type V (class-II) system, is gaining momentum because of its versatility and simplified features. These include the use of a small guide RNA devoid of trans-activating crispr RNA (tracrRNA), targeting of T-rich regions of the genome where Cas9 is not suitable for use, RNA processing capability facilitating simpler multiplexing, and its ability to generate double strand breaks (DSB) with staggered ends. Many monocot and dicot species have been successfully edited using this Cas12a system and further research is ongoing to improve its efficiency in plants, including improving the temperature stability of the Cas12a enzyme, identifying new variants of Cas12a or synthetically producing Cas12a with flexible PAM sequences. In this review we provide a comparative survey of CRISPR-Cas12a and Cas9, and provide a perspective on applications of CRISPR-Cas12 in agriculture.

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