Abstract：
:Although electroporation has been widely accepted as the main gene transfer tool, there is still considerable scope to improve the electroporation efficiency of exogenous DNAs into primary cells. Here, we developed a square-wave pulsing protocol using OptiMEM-GlutaMAX for highly efficient transfection of murine embryonic fibroblasts (MEF) and induced pluripotency stem (iPS) cells using reporter genes as well as gRNA/Cas9-encoding plasmids. An electrotransfection efficiency of > 95% was achieved for both MEF and iPS cells using reporter-encoding plasmids. The protocol was efficient for plasmid sizes ranging from 6.2 to 13.5 kb. Inducing the error prone non-homologous end joining repair by gRNA/Cas9 plasmid transfection, a high rate of targeted gene knockouts of up to 98% was produced in transgenic cells carrying a single-copy of Venus reporter. Targeted deletions in the Venus transgene were efficiently (up to 67% deletion rate) performed by co-electroporation of two gRNA-encoding plasmids. We introduced a plasmid electrotransfection protocol which is straight-forward, cost-effective, and efficient for CRISPRing murine primary cells. This protocol is promising to make targeted genetic engineering using the CRISPR/Cas9 plasmid system.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Eghbalsaied S,Hyder I,Kues WAdoi
10.1038/s41598-020-70258-wsubject
Has Abstractpub_date
2020-08-07 00:00:00pages
13332issue
1issn
2045-2322pii
10.1038/s41598-020-70258-wjournal_volume
10pub_type
杂志文章abstract：:Tumor growth and metastasis are responsible for most cancer patients' deaths. Here, we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis. Compared with proliferation, colony formation, transwell assay and transplantable tumor in nude mouse in vitr...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep05518
更新日期：2014-07-01 00:00:00
abstract：:LiYF4:Eu nanophosphors with a single tetragonal phase are synthesized, and various strategies to enhance the Eu(3+) emission from the nanophosphors are investigated. The optimized Eu(3+) concentration is 35 mol%, and the red emission peaks due to the (5)D0 →(7)FJ (J = 1 and 2) transitions of Eu(3+) ions are further en...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep07866
更新日期：2015-01-19 00:00:00
abstract：:The aim of this study was to use functional neuroimaging to investigate whether oxytocin modulates the neural response to visual food cues in brain regions involved in the control of food intake. Twenty-four normal weight volunteers received intranasal oxytocin (24 IU) or placebo in a double-blind, randomized crossove...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-04600-0
更新日期：2017-06-27 00:00:00
abstract：:DEMETER-like DNA glycosylases (DMLs) initiate the base excision repair-dependent DNA demethylation to regulate a wide range of biological processes in plants. Six putative SmDML genes, termed SmDML1-SmDML6, were identified from the genome of S. miltiorrhiza, an emerging model plant for Traditional Chinese Medicine (TC...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-25315-w
更新日期：2018-05-08 00:00:00
abstract：:Primary biliary cholangitis (PBC) is an autoimmune liver disease characterized by the destruction of interlobular biliary ductules, which progressively leads to cholestasis, hepatic fibrosis, cirrhosis, and eventually liver failure. Several mouse models have been used to clarify the pathogenesis of PBC and are general...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-15661-6
更新日期：2017-11-10 00:00:00
abstract：:Knowledge of bacteria's heat resistance is essential for developing effective thermal treatments. Choosing an appropriate test method is important to accurately determine bacteria's heat resistances. Although being a major factor to influence the thermo-tolerance of bacteria, the heating rate in samples cannot be cont...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep30758
更新日期：2016-07-28 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-67233-w
更新日期：2020-06-23 00:00:00
abstract：:Atrial fibrillation (AF) is known to cause adverse remodeling of left atrium (LA). Radiofrequency catheter ablation (RFCA) of AF is associated with decrease in LA volume. However, the impact of RFCA on left atrial appendage (LAA) volume and hemodynamic function is not fully understood. We analyzed 123 patients who und...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-26212-y
更新日期：2018-05-18 00:00:00
abstract：:The incidence of patients under 55 years old diagnosed with Prostate Cancer (EO-PCa) has increased during recent years. The molecular biology of PCa cancer in this group of patients remains unclear. Here, we applied weighted gene coexpression network analysis of the expression of miRNAs from 24 EO-PCa patients (38-45 ...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-69290-7
更新日期：2020-07-23 00:00:00
abstract：:Voltage-gated sodium (NaV) channels have been related with cell migration and invasiveness in human cancers. We previously reported the contribution of NaV1.6 channels activity with the invasion capacity of cervical cancer (CeCa) positive to Human Papilloma Virus type 16 (HPV16), which accounts for 50% of all CeCa cas...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-31364-y
更新日期：2018-08-29 00:00:00
abstract：:Delivering and expressing a gene of interest in cells or living animals has become a pivotal technique in biomedical research and gene therapy. Among viral delivery systems, adeno-associated viruses (AAVs) are relatively safe and demonstrate high gene transfer efficiency, low immunogenicity, stable long-term expressio...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-019-49624-w
更新日期：2019-09-19 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-73308-5
更新日期：2020-10-06 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep08142
更新日期：2015-01-30 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-33425-8
更新日期：2018-10-10 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-08676-6
更新日期：2017-08-18 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-05483-x
更新日期：2017-07-10 00:00:00
abstract：:Brain-computer interfaces enable active communication and execution of a pre-defined set of commands, such as typing a letter or moving a cursor. However, they have thus far not been able to infer more complex intentions or adapt more complex output based on brain signals. Here, we present neuroadaptive generative mod...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-71287-1
更新日期：2020-09-07 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep28656
更新日期：2016-06-23 00:00:00
abstract：:This study investigated the effect of various prebiotics (indigestible dextrin, α-cyclodextrin, and dextran) on human colonic microbiota at a dosage corresponding to a daily intake of 6 g of prebiotics per person (0.2% of dietary intake). We used an in vitro human colonic microbiota model based on batch fermentation s...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-18877-8
更新日期：2018-01-11 00:00:00
abstract：:The rainbow trout, a carnivorous fish, displays a 'glucose-intolerant' phenotype revealed by persistent hyperglycaemia when fed a high carbohydrate diet (HighCHO). Epigenetics refers to heritable changes in gene activity and is closely related to environmental changes and thus to metabolism adjustments governed by nut...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep32187
更新日期：2016-08-26 00:00:00
abstract：:Large-scale anthropogenic disturbances can have direct and indirect effects on marine communities, with direct effects often taking the form of widespread injury or mortality and indirect effects manifesting as changes in food web structure. Here, we report a time series that captures both direct and indirect effects ...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-62574-y
更新日期：2020-04-09 00:00:00
abstract：:Transition Metal Dichalcogenides (TMDs) are one of the most studied two-dimensional materials in the last 5-10 years due to their extremely interesting layer dependent properties. Despite the presence of vast research work on TMDs, the complex relation between the electro-chemical and physical properties make them the...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-58149-6
更新日期：2020-02-03 00:00:00
abstract：:Recent advances in genetic analyses have significantly refined human type 1 diabetes (T1D) associated loci. The goal of such effort is to identify the causal genes and have a complete understanding of the molecular pathways that independently or interactively influence cellular processes leading to the destruction of ...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-68956-6
更新日期：2020-07-21 00:00:00
abstract：:Sustainable wastewater treatment has been attracting increasing attentions over the past decades. However, the production of nitrous oxide (N2O), a potent GHG, from the energy-efficient granule-based autotrophic nitrogen removal is largely unknown. This study applied a previously established N2O model, which incorpora...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep45609
更新日期：2017-04-03 00:00:00
abstract：:Grazing and burrowing organisms usually homogenise microalgal mats that form on benthic sediments of many aquatic ecosystems. In the absence of this disruption, microalgal mats can accrete laminated deposits (stromatolites). Stromatolites are rare in modern coastal ecosystems, but persist at locations where metazoans ...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep42614
更新日期：2017-02-16 00:00:00
abstract：:We describe a new reproducibility-optimization method ROPECA for statistical analysis of proteomics data with a specific focus on the emerging data-independent acquisition (DIA) mass spectrometry technology. ROPECA optimizes the reproducibility of statistical testing on peptide-level and aggregates the peptide-level c...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-017-05949-y
更新日期：2017-07-19 00:00:00
abstract：:One of the greatest challenges in the field of biofabrication remains the discovery of suitable bioinks that satisfy physicochemical and biological requirements. Despite recent advances in tissue engineering and biofabrication, progress has been limited to the development of technologies using polymer-based materials....
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-59632-w
更新日期：2020-02-18 00:00:00
abstract：:We propose two generalized Haldane models on laser-coupling optical lattices. Laser-assisted nearest neighbour tunnelings generate artificial staggered magnetic flux, facilitating the realization of topological nontrivial band structures. As generalizations of Haldane model, these models support topological insulator ...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-30503-9
更新日期：2018-08-27 00:00:00
abstract：:Adipose stromal cells (ASC) secrete various trophic factors that assist in the protection of neurons in a variety of neuronal death models. In this study, we tested the effects of human ASC conditional medium (ASC-CM) in human amyotrophic lateral sclerosis (ALS) transgenic mouse model expressing mutant superoxide dism...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep16953
更新日期：2015-11-20 00:00:00
abstract：:The 2,5-Diketopiperazines (DKPs) constitute a large family of natural products with important biological activities. Bicyclomycin is a clinically-relevant DKP antibiotic that is the first and only member in a class known to target the bacterial transcription termination factor Rho. It derives from cyclo-(L-isoleucyl-L...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-019-56747-7
更新日期：2019-12-27 00:00:00
