scholarly journals CRISPR: Edição genômica aplicada à Oncologia / CRISPR: Genomic editing applied to Oncology

2021 ◽  
Vol 7 (9) ◽  
pp. 90908-90927
Author(s):  
Eduarda Bresolin ◽  
Paula Wiethölter ◽  
Karina Schreiner Kirsten
Keyword(s):  
Genomic Editing

scholarly journals Reversal of the Inflammatory Responses in Fabry Patient iPSC-Derived Cardiovascular Endothelial Cells by CRISPR/Cas9-Corrected Mutation

2021 ◽  
Vol 22 (5) ◽  
pp. 2381
Author(s):  
Hui-Yung Song ◽  
Yi-Ping Yang ◽  
Yueh Chien ◽  
Wei-Yi Lai ◽  
Yi-Ying Lin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Inflammatory Responses ◽  
Late Onset ◽  
Critical Role ◽  
Vascular Complications ◽  
Digital Pcr ◽  
Mapk Signaling ◽  
Autophagic Flux ◽  
Fabry Patient ◽  
Genomic Editing

The late-onset type of Fabry disease (FD) with GLA IVS4 + 919G > A mutation has been shown to lead to cardiovascular dysfunctions. In order to eliminate variations in other aspects of the genetic background, we established the isogenic control of induced pluripotent stem cells (iPSCs) for the identification of the pathogenetic factors for FD phenotypes through CRISPR/Cas9 genomic editing. We adopted droplet digital PCR (ddPCR) to efficiently capture mutational events, thus enabling isolation of the corrected FD from FD-iPSCs. Both of these exhibited the characteristics of pluripotency and phenotypic plasticity, and they can be differentiated into endothelial cells (ECs). We demonstrated the phenotypic abnormalities in FD iPSC-derived ECs (FD-ECs), including intracellular Gb3 accumulation, autophagic flux impairment, and reactive oxygen species (ROS) production, and these abnormalities were rescued in isogenic control iPSC-derived ECs (corrected FD-ECs). Microarray profiling revealed that corrected FD-derived endothelial cells reversed the enrichment of genes in the pro-inflammatory pathway and validated the downregulation of NF-κB and the MAPK signaling pathway. Our findings highlighted the critical role of ECs in FD-associated vascular dysfunctions by establishing a reliable isogenic control and providing information on potential cellular targets to reduce the morbidity and mortality of FD patients with vascular complications.

scholarly journals Epigenetic Modulation of Radiation-Induced Diacylglycerol Kinase Alpha Expression Prevents Pro-Fibrotic Fibroblast Response

Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2455
Author(s):  
Chun-Shan Liu ◽  
Reka Toth ◽  
Ali Bakr ◽  
Ashish Goyal ◽  
Md Saiful Islam ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Drug Effects ◽  
Diacylglycerol Kinase ◽  
Dna Demethylation ◽  
Dermal Fibroblasts ◽  
Common Component ◽  
Radiation Induced ◽  
Marker Expression ◽  
Epigenetic Modulation ◽  
Genomic Editing

Radiotherapy, a common component in cancer treatment, can induce adverse effects including fibrosis in co-irradiated tissues. We previously showed that differential DNA methylation at an enhancer of diacylglycerol kinase alpha (DGKA) in normal dermal fibroblasts is associated with radiation-induced fibrosis. After irradiation, the transcription factor EGR1 is induced and binds to the hypomethylated enhancer, leading to increased DGKA and pro-fibrotic marker expression. We now modulated this DGKA induction by targeted epigenomic and genomic editing of the DGKA enhancer and administering epigenetic drugs. Targeted DNA demethylation of the DGKA enhancer in HEK293T cells resulted in enrichment of enhancer-related histone activation marks and radiation-induced DGKA expression. Mutations of the EGR1-binding motifs decreased radiation-induced DGKA expression in BJ fibroblasts and caused dysregulation of multiple fibrosis-related pathways. EZH2 inhibitors (GSK126, EPZ6438) did not change radiation-induced DGKA increase. Bromodomain inhibitors (CBP30, JQ1) suppressed radiation-induced DGKA and pro-fibrotic marker expression. Similar drug effects were observed in donor-derived fibroblasts with low DNA methylation. Overall, epigenomic manipulation of DGKA expression may offer novel options for a personalized treatment to prevent or attenuate radiotherapy-induced fibrosis.

scholarly journals Non-viral delivery of CRISPR–Cas9 complexes for targeted gene editing via a polymer delivery system

Gene Therapy ◽  
2021 ◽  
Author(s):  
Jonathan O’Keeffe Ahern ◽  
Irene Lara-Sáez ◽  
Dezhong Zhou ◽  
Rodolfo Murillas ◽  
Jose Bonafont ◽  
...  
Keyword(s):  
Delivery System ◽  
Gene Editing ◽  
Transfection Efficiency ◽  
Attractive Alternative ◽  
Safe Delivery ◽  
Guide Rna ◽  
Recessive Dystrophic Epidermolysis Bullosa ◽  
Targeted Gene Editing ◽  
Genomic Editing ◽  
Polymeric Vectors

AbstractRecent advances in molecular biology have led to the CRISPR revolution, but the lack of an efficient and safe delivery system into cells and tissues continues to hinder clinical translation of CRISPR approaches. Polymeric vectors offer an attractive alternative to viruses as delivery vectors due to their large packaging capacity and safety profile. In this paper, we have demonstrated the potential use of a highly branched poly(β-amino ester) polymer, HPAE-EB, to enable genomic editing via CRISPRCas9-targeted genomic excision of exon 80 in the COL7A1 gene, through a dual-guide RNA sequence system. The biophysical properties of HPAE-EB were screened in a human embryonic 293 cell line (HEK293), to elucidate optimal conditions for efficient and cytocompatible delivery of a DNA construct encoding Cas9 along with two RNA guides, obtaining 15–20% target genomic excision. When translated to human recessive dystrophic epidermolysis bullosa (RDEB) keratinocytes, transfection efficiency and targeted genomic excision dropped. However, upon delivery of CRISPR–Cas9 as a ribonucleoprotein complex, targeted genomic deletion of exon 80 was increased to over 40%. Our study provides renewed perspective for the further development of polymer delivery systems for application in the gene editing field in general, and specifically for the treatment of RDEB.

scholarly journals Precise genomic editing

Nature ◽  
2013 ◽  
Vol 493 (7431) ◽  
pp. 137-137
Keyword(s):  
Genomic Editing

Legal support for the creation and use of genetic technologies in medicine

2020 ◽  
pp. 77-78
Author(s):  
В.В. Лапаева
Keyword(s):  
Human Rights ◽  
Great Britain ◽  
Human Genome ◽  
Germ Line ◽  
Genetic Integrity ◽  
Scientific Creativity ◽  
Genetic Technologies ◽  
Genomic Editing ◽  
The Given ◽  
Proper Balance

Правовая политика России в сфере создания и применения генетических технологий в медицине не обеспечивает в должной мере баланс между системой прав человека, гарантирующих защиту его достоинства и охрану здоровья, и свободой научного творчества. Несогласованность этих прав человека проявляется в том, что при отсутствии запретов на исследования с применением технологий наследуемого редактирования генома человека законодательство лишает патентоспособности любые технологии по модификации генетической целостности клеток зародышевой линии человека. Последовательный правовой подход предполагает введение ограничений на возможность геномного редактирования зародышевой линии и запрета на патентование способов такого редактирования, не выходящего за рамки заданных ограничений. При разработке такого похода целесообразно учесть опыт Великобритании. The Russian legal policy in the field of the medicine genetic technologies creation and application does not provide a proper balance between the system of human rights that guarantee the protection of its dignity and health, and the freedom of scientific creativity. The non-coordination of these human rights is manifested in the fact that in the absence of prohibitions on research using technologies of human genome inherited editing, the law deprives patentability of any technology for modifying the genetic integrity of human germline cells. A consistent legal approach involves the restrictions on the possibility of the germ line genomic editing and the prohibition of patenting the methods of such editing, which does not go beyond the given restrictions. It is advisable to take into account the experience of Great Britain.

scholarly journals 25 A ZEBRAFISH MODEL OF MYELODYSPLASTIC SYNDROME PRODUCED THROUGH TET2 GENOMIC EDITING

2015 ◽  
Vol 39 ◽  
pp. S11
Author(s):  
E. Gjini ◽  
M.A.R.C. Mansour ◽  
S. He ◽  
A. Nguyen ◽  
M. Ko ◽  
...  
Keyword(s):  
Myelodysplastic Syndrome ◽  
Zebrafish Model ◽  
Genomic Editing

scholarly journals Differential enrichment of yeast DNA in SARS-CoV-2 and related genomes supports synthetic origin hypothesis

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 912
Author(s):  
Andreas Martin Lisewski
Keyword(s):  
Large Scale ◽  
The Other ◽  
Yeast Cells ◽  
Virus Family ◽  
Spike Gene ◽  
Viral Replicase ◽  
Genomic Editing ◽  
Chromosomal Sequences ◽  
Bat Coronavirus ◽  
Origin Hypothesis

Background: Knowledge about the origin of SARS-CoV-2 is necessary for both a biological and epidemiological understanding of the COVID-19 pandemic. Evidence suggests that a proximal evolutionary ancestor of SARS-CoV-2 belongs to the bat coronavirus family. However, as further evidence for a direct zoonosis remains limited, alternative modes of SARS-CoV-2 biogenesis should be also considered.    Results: Here we show that the genomes from SARS-CoV-2 and from SARS-CoV-1 are differentially enriched with short chromosomal sequences from the yeast S. cerevisiae at focal positions that are known to be critical for virus replication, host cell invasion, and host immune response. Specifically, for SARS-CoV-2, we identify two sites: one at the start of the viral replicase domain, and the other at the end of the spike gene past its critical domain junction; for SARS-CoV-1, one at the start of the RNA dependent RNA polymerase gene, and the other at the start of the spike protein’s receptor binding domain. As yeast is not a natural host for this virus family, we propose a directed passage model for viral constructs, including virus replicase, in yeast cells based on co-transformation of virus DNA plasmids carrying yeast selectable genetic markers followed by intra-chromosomal homologous recombination through gene conversion. Highly differential sequence homology data across yeast chromosomes congruent with chromosomes harboring specific auxotrophic markers further support this passage model. Model and data together allow us to infer a hypothetical tripartite genome assembly scheme for the synthetic biogenesis of SARS-CoV-2 and SARS-CoV-1.   Conclusions: These results provide evidence that the genome sequences of SARS-CoV-1, SARS-CoV-2, but not that of RaTG13, BANAL-20-52 and all other closest SARS coronavirus family members identified, are carriers of distinct homology signals that might point to large-scale genomic editing during a passage of directed replication and chromosomal integration inside genetically modified yeast cells.

scholarly journals New genome editing technologies in the treatment of X-linked adrenoleukodystrophy

2020 ◽  
Vol 65 (2) ◽  
pp. 104-107
Author(s):  
V. Yu. Voinova ◽  
M. А. Shkolnikova ◽  
E. A. Nikolaeva
Keyword(s):  
Lentiviral Vector ◽  
Allogeneic Transplantation ◽  
Graft Versus Host Reaction ◽  
Gene Encoding ◽  
Graft Versus Host ◽  
Alternative Approach ◽  
Cd34 Cells ◽  
Male Patients ◽  
Genomic Editing ◽  
Type Sequence

X-linked adrenoleukodystrophy is a severe progressive neurological disease that is predominantly found in male patients and caused by mutations in the X-linked ABCD1 gene encoding peroxisome transport protein. The disease is clinically characterized by two main phenotypes: the most severe infant cerebral form and adrenomyeloneuropathy. The disease is treated by allogeneic transplantation of hematopoietic cells from a healthy donor to stop progression, and gene therapy with a self-activating lentiviral vector, the carrier of the functional gene ABCD1. Each method has its own limitations. The authors present and theoretically substantiate an alternative approach to the treatment of adrenoleukodystrophy; they propose to modify the autologous CD34+ cells from the patient using genomic editing, in order to replace the mutant DNA sequence of ABCD1 gene with a wild-type sequence, while replacing the mutant protein in the edited cells. The edited autologous CD34+ cells can be introduced by their transplantation into the bone marrow or by a series of repeated intravenous infusions. This method will allow avoiding both the search for a donor and the graft-versus-host reaction

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