scholarly journals Translational Read-Through Therapy of RPGR Nonsense Mutations

2020 ◽  
Vol 21 (22) ◽  
pp. 8418
Author(s):  
Christine Vössing ◽  
Marta Owczarek-Lipska ◽  
Kerstin Nagel-Wolfrum ◽  
Charlotte Reiff ◽  
Christoph Jüschke ◽  
...  
Keyword(s):  
Retinitis Pigmentosa ◽  
Western Blot ◽  
Primary Cilium ◽  
Genetic Diseases ◽  
The Novel ◽  
Rt Pcr ◽  
Nonsense Mutations ◽  
Premature Termination Codons ◽  
New Treatment ◽  
First Time

X-chromosomal retinitis pigmentosa (RP) frequently is caused by mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. We evaluated the potential of PTC124 (Ataluren, TranslamaTM) treatment to promote ribosomal read-through of premature termination codons (PTC) in RPGR. Expression constructs in HEK293T cells showed that the efficacy of read-through reagents is higher for UGA than UAA PTCs. We identified the novel hemizygous nonsense mutation c.1154T > A, p.Leu385* (NM_000328.3) causing a UAA PTC in RPGR and generated patient-derived fibroblasts. Immunocytochemistry of serum-starved control fibroblasts showed the RPGR protein in a dot-like expression pattern along the primary cilium. In contrast, RPGR was no longer detectable at the primary cilium in patient-derived cells. Applying PTC124 restored RPGR at the cilium in approximately 8% of patient-derived cells. RT-PCR and Western blot assays verified the pathogenic mechanisms underlying the nonsense variant. Immunofluorescence stainings confirmed the successful PTC124 treatment. Our results showed for the first time that PTC124 induces read-through of PTCs in RPGR and restores the localization of the RPGR protein at the primary cilium in patient-derived cells. These results may provide a promising new treatment option for patients suffering from nonsense mutations in RPGR or other genetic diseases.

scholarly journals Efficient Suppression of Endogenous CFTR Nonsense Mutations Using Anticodon Engineered Transfer RNAs

2021 ◽  
Author(s):  
Wooree Ko ◽  
Joseph J. Porter ◽  
Matthew T. Sipple ◽  
Katherine M. Edwards ◽  
John D. Lueck
Keyword(s):  
Cystic Fibrosis ◽  
Mammalian Cells ◽  
Genetic Diseases ◽  
Protein Translation ◽  
Nonsense Mutations ◽  
Channel Function ◽  
Associated Diseases ◽  
Highly Active ◽  
Premature Termination Codons ◽  
Efficient Suppression

Nonsense mutations or premature termination codons (PTCs) comprise ~11% of all genetic lesions, which result in over 7,000 distinct genetic diseases. Due to their outsized impact on human health, considerable effort has been made to find therapies for nonsense-associated diseases. Suppressor tRNAs have long been identified as a possible therapeutic for nonsense-associated diseases, however their ability to inhibit nonsense-mediated mRNA decay (NMD) and support significant protein translation from endogenous transcripts has not been determined in mammalian cells. Here we investigated the ability of anticodon edited (ACE)-tRNAs to suppress cystic fibrosis (CF) causing PTCs in the cystic fibrosis transmembrane regulator (CFTR) gene in gene-edited immortalized human bronchial epithelial (16HBEge) cells. Delivery of ACE-tRNAs to 16HBEge cells harboring three common CF mutations G542X-, R1162X- and W1282X-CFTR PTCs significantly inhibited NMD and rescued endogenous mRNA expression. Furthermore, delivery of our highly active leucine encoding ACE-tRNA resulted in rescue of W1282X-CFTR channel function to levels that significantly exceed the necessary CFTR channel function for therapeutic relevance. This study establishes the ACE-tRNA approach as a potential stand-alone therapeutic for nonsense-associated diseases due to its ability to rescue both mRNA and full-length protein expression from PTC containing endogenous genes.

scholarly journals 2-Guanidino-quinazoline promotes the readthrough of nonsense mutations underlying human genetic diseases

2021 ◽  
Author(s):  
Laure Bidou ◽  
Olivier Bugaud ◽  
Goulven Merer ◽  
Matthieu Coupet ◽  
Isabelle Hatin ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
High Throughput Screening ◽  
Therapeutic Strategy ◽  
Genetic Diseases ◽  
Medical Applications ◽  
Reporter Cell Line ◽  
Reporter Cell ◽  
Nonsense Mutations ◽  
Premature Termination Codons ◽  
Clinical Potential

Premature termination codons (PTCs) account for 10% to 20% of genetic diseases in humans. The gene inactivation resulting from PTC can be counteracted by the use of drugs stimulating PTC readthrough, thereby restoring production of the full-length protein. However, a greater chemical variety of readthrough inducers is required to broaden the medical applications of this therapeutic strategy. In this study, we developed a new reporter cell line and performed high-throughput screening (HTS) to identify potential new readthrough inducers. After three successive assays, we isolated 2-guanidino-quinazoline (TLN468). We assessed the clinical potential of this drug as a potent readthrough inducer on the 40 PTCs most frequently responsible for Duchenne muscular dystrophy. We found that TLN468 was more efficient than gentamicin, and acted on a broader range of sequences, without inducing the readthrough of natural stop codons.

scholarly journals Strategies against Nonsense: Oxadiazoles as Translational Readthrough-Inducing Drugs (TRIDs)

2019 ◽  
Vol 20 (13) ◽  
pp. 3329 ◽  
Author(s):  
Ambra Campofelice ◽  
Laura Lentini ◽  
Aldo Di Leonardo ◽  
Raffaella Melfi ◽  
Marco Tutone ◽  
...  
Keyword(s):  
Small Molecules ◽  
Genetic Diseases ◽  
Nonsense Mutations ◽  
New Class ◽  
Premature Termination Codons ◽  
Translational Readthrough ◽  
Action Strategies ◽  
Oxadiazole Derivatives ◽  
Cftr Protein ◽  
Cystic Fibrosis Transmembrane

This review focuses on the use of oxadiazoles as translational readthrough-inducing drugs (TRIDs) to rescue the functional full-length protein expression in mendelian genetic diseases caused by nonsense mutations. These mutations in specific genes generate premature termination codons (PTCs) responsible for the translation of truncated proteins. After a brief introduction on nonsense mutations and their pathological effects, the features of various classes of TRIDs will be described discussing differences or similarities in their mechanisms of action. Strategies to correct the PTCs will be presented, particularly focusing on a new class of Ataluren-like oxadiazole derivatives in comparison to aminoglycosides. Additionally, recent results on the efficiency of new candidate TRIDs in restoring the production of the cystic fibrosis transmembrane regulator (CFTR) protein will be presented. Finally, a prospectus on complementary strategies to enhance the effect of TRIDs will be illustrated together with a conclusive paragraph about perspectives, opportunities, and caveats in developing small molecules as TRIDs.

Buried Narratives: Exhuming the Mother's Story in Oliver Twist

2018 ◽  
Vol 8 (3) ◽  
pp. 247-266
Author(s):  
Michelle L. Wilson
Keyword(s):  
Social Change ◽  
Maternal Inheritance ◽  
The Novel ◽  
Poor Law ◽  
The Family ◽  
The Law ◽  
First Time ◽  
Critical Consideration

Initially, Oliver Twist (1839) might seem representative of the archetypal male social plot, following an orphan and finding him a place by discovering the father and settling the boy within his inheritance. But Agnes Fleming haunts this narrative, undoing its neat, linear transmission. This reconsideration of maternal inheritance and plot in the novel occurs against the backdrop of legal and social change. I extend the critical consideration of the novel's relationship to the New Poor Law by thinking about its reflection on the bastardy clauses. And here, of course, is where the mother enters. Under the bastardy clauses, the responsibility for economic maintenance of bastard children was, for the first time, legally assigned to the mother, relieving the father of any and all obligation. Oliver Twist manages to critique the bastardy clauses for their release of the father, while simultaneously embracing the placement of the mother at the head of the family line. Both Oliver and the novel thus suggest that it is the mother's story that matters, her name through which we find our own. And by containing both plots – that of the father and the mother – Oliver Twist reveals the violence implicit in traditional modes of inheritance in the novel and under the law.

Direct Read and Quantify Damage Nucleotide from an Oligonucleotide Using a Single Molecule Interface

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Meng-Yin Li ◽  
Jie Yang ◽  
Ya-Qian Wang ◽  
Xue-Yuan Wu ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Single Molecule ◽  
Genetic Diseases ◽  
High Sensitivity ◽  
Dna Lesions ◽  
Lesion Detection ◽  
The Novel ◽  
Structural Variations ◽  
Dna Lesion ◽  
Base Lesion

DNA lesion such as metholcytosine(<sup>m</sup>C), 8-OXO-guanine(<sup>O</sup>G), inosine(I) <i>etc</i> could cause the genetic diseases. Identification of the varieties of lesion bases are usually beyond the capability of conventional DNA sequencing which is mainly designed to discriminate four bases only. Therefore, lesion detection remain challenge due to the massive varieties and less distinguishable readouts for minor structural variations. Moreover, standard amplification and labelling hardly works in DNA lesions detection. Herein, we designed a single molecule interface from the mutant K238Q Aerolysin, whose confined sensing region shows the high compatible to capture and then directly convert each base lesion into distinguishable current readouts. Compared with previous single molecule sensing interface, the resolution of the K238Q Aerolysin nanopore is enhanced by 2-order. The novel K238Q could direct discriminate at least 3 types (<sup>m</sup>C, <sup>O</sup>G, I) lesions without lableing and quantify modification sites under mixed hetero-composition condition of oligonucleotide. Such nanopore could be further applied to diagnose genetic diseases at high sensitivity.

Application of Novel Low Current OBIRCH Amplifier and Nanoprobing to Identify Subtle Leakages in Advanced Node Technologies

2018 ◽  
Author(s):  
Satish Kodali ◽  
Liangshan Chen ◽  
Yuting Wei ◽  
Tanya Schaeffer ◽  
Chong Khiam Oh
Keyword(s):  
Signal To Noise Ratio ◽  
Semiconductor Industry ◽  
Fault Isolation ◽  
Ring Oscillator ◽  
High Signal ◽  
The Novel ◽  
Resistance Change ◽  
Three Samples ◽  
Relative Threshold ◽  
First Time

Abstract Optical beam induced resistance change (OBIRCH) is a very well-adapted technique for static fault isolation in the semiconductor industry. Novel low current OBIRCH amplifier is used to facilitate safe test condition requirements for advanced nodes. This paper shows the differences between the earlier and novel generation OBIRCH amplifiers. Ring oscillator high standby leakage samples are analyzed using the novel generation amplifier. High signal to noise ratio at applied low bias and current levels on device under test are shown on various samples. Further, a metric to demonstrate the SNR to device performance is also discussed. OBIRCH analysis is performed on all the three samples for nanoprobing of, and physical characterization on, the leakage. The resulting spots were calibrated and classified. It is noted that the calibration metric can be successfully used for the first time to estimate the relative threshold voltage of individual transistors in advanced process nodes.

Rhythmic Prose in Imperial Greek Literature

2018 ◽  
Author(s):  
G. O. Hutchinson
Keyword(s):  
The Novel ◽  
Greek Literature ◽  
System A ◽  
First Time

The chapter looks at the division between poetry and prose in ancient and other literatures, and shows the importance of rhythmic patterning in ancient prose. The development of rhythmic prose in Greek and Latin is sketched, the system explained and illustrated (from Latin). It is firmly established, for the first time, which of the main Greek non-Christian authors 31 BC–AD 300 write rhythmically. The method takes a substantial sample of random sentence-endings (usually 400) from each of a large number of Imperial authors; it compares that sample with one sample of the same size (400) drawn randomly from a range of authors earlier than the invention of this rhythmic system. A particular sort of X2-test is applied. Many Imperial authors, it emerges, write rhythmically; many do not. The genres most likely to offer rhythmic writing are, unexpectedly, narrative: historiography and the novel.

Kidnapped

2014 ◽  
Author(s):  
Robert Louis Stevenson ◽  
Ian Duncan
Keyword(s):  
West Coast ◽  
Historical Novel ◽  
The Novel ◽  
The West ◽  
Scottish Highlands ◽  
Cross Country ◽  
First Time ◽  
Adventure Stories

Your bed shall be the moorcock’s, and your life shall be like the hunted deer’s, and ye shall sleep with your hand upon your weapons.’ Tricked out of his inheritance, shanghaied, shipwrecked off the west coast of Scotland, David Balfour finds himself fleeing for his life in the dangerous company of Jacobite outlaw and suspected assassin Alan Breck Stewart. Their unlikely friendship is put to the test as they dodge government troops across the Scottish Highlands. Set in the aftermath of the 1745 rebellion, Kidnapped transforms the Romantic historical novel into the modern thriller. Its heart-stopping scenes of cross-country pursuit, distilled to a pure intensity in Stevenson’s prose, have become a staple of adventure stories from John Buchan to Alfred Hitchcock and Ian Fleming. Kidnapped remains as exhilarating today as when it was first published in 1886. This new edition is based on the 1895 text, incorporating Stevenson’s last thoughts about the novel before his death. It includes Stevenson’s ‘Note to Kidnapped’, reprinted for the first time since 1922.

scholarly journals Phylogenomic Evidence of Reinfection and Persistence of SARS-CoV-2: First Report from Colombia

Vaccines ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 282
Author(s):  
Juan David Ramírez ◽  
Marina Muñoz ◽  
Nathalia Ballesteros ◽  
Luz H. Patiño ◽  
Sergio Castañeda ◽  
...  
Keyword(s):  
Viral Persistence ◽  
Transmission Dynamics ◽  
Polymorphism Analysis ◽  
Rt Pcr ◽  
Paired Samples ◽  
Infection Dynamics ◽  
Oxford Nanopore ◽  
Novel Variants ◽  
First Time ◽  
Oxford Nanopore Technologies

The continuing evolution of SARS-CoV-2 and the emergence of novel variants have raised concerns about possible reinfection events and potential changes in the coronavirus disease 2019 (COVID-19) transmission dynamics. Utilizing Oxford Nanopore technologies, we sequenced paired samples of three patients with positive RT-PCR results in a 1–2-month window period, and subsequent phylogenetics and genetic polymorphism analysis of these genomes was performed. Herein, we report, for the first time, genomic evidence of one case of reinfection in Colombia, exhibiting different SARS-CoV-2 lineage classifications between samples (B.1 and B.1.1.269). Furthermore, we report two cases of possible viral persistence, highlighting the importance of deepening our understanding on the evolutionary intra-host traits of this virus throughout different timeframes of disease progression. These results emphasize the relevance of genomic surveillance as a tool for understanding SARS-CoV-2 infection dynamics, and how this may translate effectively to future control and mitigations efforts, such as the national vaccination program.

scholarly journals Antimicrobial Activity of L-Lysine and Poly-L-Lysine with Pulsed Electric Fields

2021 ◽  
Vol 11 (6) ◽  
pp. 2708
Author(s):  
Jurgita Švedienė ◽  
Vitalij Novickij ◽  
Rokas Žalnėravičius ◽  
Vita Raudonienė ◽  
Svetlana Markovskaja ◽  
...  
Keyword(s):  
Electric Fields ◽  
Pulsed Electric Fields ◽  
Antimicrobial Treatment ◽  
Treatment Development ◽  
E Coli ◽  
Yeast Fungi ◽  
New Treatment ◽  
First Time ◽  
Resistant Microorganism ◽  
Susceptibility Patterns

For the first time, the possibility to use L-lysine (Lys) and poly-L-lysine (PLL) as additives with pulsed electric fields (PEF) for antimicrobial treatment is reported. The antimicrobial efficacy of Lys and PLL for Escherichia coli, Staphylococcus aureus, Trichophyton rubrum and Candida albicans was determined. Inactivation of microorganisms was also studied by combining Lys and PLL with PEF of 15 and 30 kV/cm. For PEF treatment, pulses of 0.5, 1, 10 or 100 μs were applied in a sequence of 10 to 5000 at 1 kHz frequency. The obtained results showed that 100 μs pulses were the most effective in combination with Lys and PLL for all microorganisms. Equivalent energy PEF bursts with a shorter duration of the pulse were less effective independently on PEF amplitude. Additionally, various treatment susceptibility patterns of microorganisms were determined and reported. In this study, the Gram-negative E. coli was the most treatment-resistant microorganism. Nevertheless, inactivation rates exceeding 2 log viability reduction were achieved for all analyzed yeast, fungi, and bacteria. This methodology could be used for drug-resistant microorganism’s new treatment development.

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