scholarly journals CRISPR-RT: A web service for designing CRISPR-C2c2 crRNA with improved target specificity

2017 ◽  
Author(s):  
Houxiang Zhu ◽  
Emily Richmond ◽  
Chun Liang
Keyword(s):  
Web Service ◽  
Genome Editing ◽  
Rna Editing ◽  
Model Organisms ◽  
Future Research ◽  
Target Specificity ◽  
Link Type ◽  
Wide Range ◽  
Set Up

AbstractCRISPR-Cas systems have been successfully applied in genome editing. Recently, the CRISPR-C2c2 system has been reported as a tool for RNA editing. Here we describe CRISPR-RT (CRISPR RNA-Targeting), the first web service to help biologists design the crRNA with improved target specificity for the CRISPR-C2c2 system. CRISPR-RT allows users to set up a wide range of parameters, making it highly flexible for current and future research in CRISPR-based RNA editing. CRISPR-RT covers major model organisms and can be easily extended to cover other species. CRISPR-RT will empower researchers in RNA editing. It is available at http://bioinfolab.miamioh.edu/CRISPR-RT.

A Review of Theoretical and Experimental Research on Various Autofrettage Processes

2018 ◽  
Vol 140 (5) ◽  
Author(s):  
Rajkumar Shufen ◽  
Uday S. Dixit
Keyword(s):  
Experimental Studies ◽  
Theoretical Models ◽  
Pressure Vessels ◽  
Future Research ◽  
Spherical Vessel ◽  
Considerable Research ◽  
Wide Range ◽  
Set Up ◽  
Compressive Residual Stresses ◽  
Spherical Pressure

Autofrettage is a metal forming technique widely incorporated for strengthening the thick-walled cylindrical and spherical pressure vessels. The technique is based on the principle of initially subjecting the cylindrical or spherical vessel to partial plastic deformation and then unloading it; as a result of which compressive residual stresses are set up. On the basis of the type of the forming load, autofrettage can be classified into hydraulic, swage, explosive, thermal, and rotational. Considerable research studies have been carried out on autofrettage with a variety of theoretical models and experimental methods. This paper presents an extensive review of various types of autofrettage processes. A wide range of theoretical models and experimental studies are described. Optimization of an autofrettage process is also discussed. Based on the review, some challenging issues and key areas for future research are identified.

scholarly journals PinAPL-Py: A comprehensive web-application for the analysis of CRISPR/Cas9 screens

2017 ◽  
Author(s):  
Philipp N. Spahn ◽  
Tyler Bath ◽  
Ryan J. Weiss ◽  
Jihoon Kim ◽  
Jeffrey D. Esko ◽  
...  
Keyword(s):  
Web Application ◽  
Large Scale ◽  
Sequencing Data ◽  
Bioinformatic Tools ◽  
Link Type ◽  
Screening Experiments ◽  
Independent Analysis ◽  
Wide Range ◽  
Set Up ◽  
Sequence Quality

AbstractBackgroundLarge-scale genetic screens using CRISPR/Cas9 technology have emerged as a major tool for functional genomics. With its increased popularity, experimental biologists frequently acquire large sequencing datasets for which they often do not have an easy analysis option. While a few bioinformatic tools have been developed for this purpose, their utility is still hindered either due to limited functionality or the requirement of bioinformatic expertise.ResultsTo make sequencing data analysis of CRISPR/Cas9 screens more accessible to a wide range of scientists, we developed a Platform-independent Analysis of Pooled Screens using Python (PinAPL-Py), which is operated as an intuitive web-service. PinAPL-Py implements state-of-the-art tools and statistical models, assembled in a comprehensive workflow covering sequence quality control, automated sgRNA sequence extraction, alignment, sgRNA enrichment/depletion analysis and gene ranking. The workflow is set up to use a variety of popular sgRNA libraries as well as custom libraries that can be easily uploaded. Various analysis options are offered, suitable to analyze a large variety of CRISPR/Cas9 screening experiments. Analysis output includes ranked lists of sgRNAs and genes, and publication-ready plots.ConclusionsPinAPL-Py helps to advance genome-wide screening efforts by combining comprehensive functionality with user-friendly implementation. PinAPL-Py is freely accessible at http://pinapl-py.ucsd.edu with instructions, documentation and test datasets. The source code is available at https://github.com/LewisLabUCSD/PinAPL-Py

scholarly journals SNP-CRISPR: a web tool for SNP-specific genome editing

2019 ◽  
Author(s):  
Chiao-Lin Chen ◽  
Jonathan Rodiger ◽  
Verena Chung ◽  
Raghuvir Viswanatha ◽  
Stephanie E. Mohr ◽  
...  
Keyword(s):  
Genome Editing ◽  
Model Systems ◽  
Model Organisms ◽  
Data Sets ◽  
Nucleotide Polymorphisms ◽  
Web Tool ◽  
Guide Rna ◽  
Wide Range ◽  
Sgrna Design ◽  
Reference Genomes

ABSTRACTCRISPR-Cas9 is a powerful genome editing technology in which a single guide RNA (sgRNA) confers target site specificity to achieve Cas9-mediated genome editing. Numerous sgRNA design tools have been developed based on reference genomes for humans and model organisms. However, existing resources are not optimal as genetic mutations or single nucleotide polymorphisms (SNPs) within the targeting region affect the efficiency of CRISPR-based approaches by interfering with guide-target complementarity. To facilitate identification of sgRNAs (1) in non-reference genomes, (2) across varying genetic backgrounds, or (3) for specific targeting of SNP-containing alleles, for example, disease relevant mutations, we developed a web tool, SNP-CRISPR (https://www.flyrnai.org/tools/snp_crispr/). SNP-CRISPR can be used to design sgRNAs based on public variant data sets or user-identified variants. In addition, the tool computes efficiency and specificity scores for sgRNA designs targeting both the variant and the reference. Moreover, SNP-CRISPR provides the option to upload multiple SNPs and target single or multiple nearby base changes simultaneously with a single sgRNA design. Given these capabilities, SNP-CRISPR has a wide range of potential research applications in model systems and for design of sgRNAs for disease-associated variant correction.

scholarly journals Evolutionary Analyses of RNA Editing and Amino Acid Recoding in Cephalopods

2019 ◽  
Author(s):  
Mingye (Christina) Wang ◽  
Erik Mohlhenrich
Keyword(s):  
Amino Acid ◽  
Candidate Genes ◽  
Rna Editing ◽  
Amino Acid Level ◽  
Nucleotide Composition ◽  
Model Organisms ◽  
Future Research ◽  
Amino Acid Substitutions ◽  
Behavioral Complexity ◽  
Fitness Advantage

AbstractRNA editing is a post-transcriptional modification process that alters nucleotides of mRNA and consequently the amino acids of the translated protein without changing the original DNA sequence. In human and other mammals, amino acid recoding from RNA editing is rare, and most edits are non-adaptive and provide no fitness advantage (1). However, recently it was discovered that in soft-bodied cephalopods, which are exceptionally intelligent and include squid, octopus, and cuttlefish, RNA editing is widespread and positively selected (2). To examine the effects of RNA editing on individual genes, we developed a “diversity score” system that quantitatively assesses the amount of diversity generated in each gene, incorporating combinatorial diversity and the radicalness of amino acid changes. Using this metric, we compiled a list of top 100 genes across the cephalopod species that are most diversified by RNA editing. This list of candidate genes provides directions for future research into the specific functional impact of RNA editing in terms of protein structure and cellular function on individual proteins. Additionally, considering the connection of RNA editing to the nervous system, and the exceptional intelligence of cephalopod, the candidate genes may shed light to the molecular development of behavioral complexity and intelligence. To further investigate global influences of RNA editing on the transcriptome, we investigated changes in nucleotide composition and codon usage biases in edited genes and coleoid transcriptome in general. Results show that these features indeed correlate with editing and may correspond to causes or effects of RNA editing. In addition, we characterized the unusual RNA editing in cephalopods by analyzing ratio of radical to conservative amino acid substitutions (R/C) and distribution of amino acid recoding from editing. Our results show that compared to model organisms, editing in cephalopods have significantly decreased R/C ratio and distinct distribution of amino acid substitutions that favor conservative over radical changes, indicating selection at the amino acid level and providing a potential mechanism for the evolution of widespread RNA editing in cephalopods.

scholarly journals SNP-CRISPR: A Web Tool for SNP-Specific Genome Editing

2019 ◽  
Vol 10 (2) ◽  
pp. 489-494 ◽  
Author(s):  
Chiao-Lin Chen ◽  
Jonathan Rodiger ◽  
Verena Chung ◽  
Raghuvir Viswanatha ◽  
Stephanie E. Mohr ◽  
...  
Keyword(s):  
Genome Editing ◽  
Model Systems ◽  
Model Organisms ◽  
Data Sets ◽  
Nucleotide Polymorphisms ◽  
Web Tool ◽  
Guide Rna ◽  
Wide Range ◽  
Sgrna Design ◽  
Reference Genomes

CRISPR-Cas9 is a powerful genome editing technology in which a single guide RNA (sgRNA) confers target site specificity to achieve Cas9-mediated genome editing. Numerous sgRNA design tools have been developed based on reference genomes for humans and model organisms. However, existing resources are not optimal as genetic mutations or single nucleotide polymorphisms (SNPs) within the targeting region affect the efficiency of CRISPR-based approaches by interfering with guide-target complementarity. To facilitate identification of sgRNAs (1) in non-reference genomes, (2) across varying genetic backgrounds, or (3) for specific targeting of SNP-containing alleles, for example, disease relevant mutations, we developed a web tool, SNP-CRISPR (https://www.flyrnai.org/tools/snp_crispr/). SNP-CRISPR can be used to design sgRNAs based on public variant data sets or user-identified variants. In addition, the tool computes efficiency and specificity scores for sgRNA designs targeting both the variant and the reference. Moreover, SNP-CRISPR provides the option to upload multiple SNPs and target single or multiple nearby base changes simultaneously with a single sgRNA design. Given these capabilities, SNP-CRISPR has a wide range of potential research applications in model systems and for design of sgRNAs for disease-associated variant correction.

scholarly journals CRISPR-DT: designing gRNAs for the CRISPR-Cpf1 system with improved target efficiency and specificity

2018 ◽  
Author(s):  
Houxiang Zhu ◽  
Chun Liang
Keyword(s):  
Machine Learning ◽  
Web Service ◽  
Genome Editing ◽  
Structural Features ◽  
Learning Technology ◽  
Dna Targeting ◽  
Link Type ◽  
Svm Model ◽  
Service Application

AbstractThe CRISPR-Cpf1 system has been successfully applied in genome editing. However, target efficiency of the CRISPR-Cpf1 system varies among different gRNA sequences. We reanalyzed the published CRISPR-Cpf1 gRNAs data and found many sequence and structural features related to their target efficiency. Using machine learning technology, a SVM model was created to predict target efficiency for any given gRNAs. We have developed the first web service application, CRISPR-DT (CRISPR DNA Targeting), to help users design optimal gRNAs for the CRISPR-Cpf1 system by considering both target efficiency and specificity. CRISPR-DT is available at http://bioinfolab.miamioh.edu/CRISPR-DT.

scholarly journals A novel workflow to improve multi-locus genotyping of wildlife species: an experimental set-up with a known model system

2019 ◽  
Author(s):  
Mark A.F. Gillingham ◽  
B. Karina Montero ◽  
Kerstin Wihelm ◽  
Kara Grudzus ◽  
Simone Sommer ◽  
...  
Keyword(s):  
Sequence Data ◽  
Model Organism ◽  
Model Systems ◽  
Model Organisms ◽  
Amplification Efficiency ◽  
Amplification Bias ◽  
Link Type ◽  
Multiple Loci ◽  
Number Variation ◽  
Set Up

ABSTRACTGenotyping novel complex multigene systems is particularly challenging in non-model organisms. Target primers frequently amplify simultaneously multiple loci leading to high PCR and sequencing artefacts such as chimeras and allele amplification bias. Most next-generation sequencing genotyping pipelines have been validated in non-model systems whereby the real genotype is unknown and the generation of artefacts may be highly repeatable. Further hindering accurate genotyping, the relationship between artefacts and copy number variation (CNV) within a PCR remains poorly described. Here we investigate the latter by experimentally combining multiple known major histocompatibility complex (MHC) haplotypes of a model organism (chicken, Gallus gallus, 43 artificial genotypes with 2-13 alleles per amplicon). In addition to well defined “optimal” primers, we simulated a non-model species situation by designing “naive” primers, with sequence data from closely related Galliform species. We applied a novel open-source genotyping pipeline (ACACIA) to the data, and compared its performance with another, previously published, pipeline. ACACIA yielded very high allele calling accuracy (>98%). Non-chimeric artefacts increased linearly with increasing CNV but chimeric artefacts leveled when amplifying more than 4-6 alleles. As expected, we found heterogeneous amplification efficiency of allelic variants when co-amplifying multiple loci. Using our validated ACACIA pipeline and the example data of this study, we discuss in detail the pitfalls researchers should avoid in order to reliably genotype complex multigene systems. ACACIA and the datasets used in this study are publicly available at GitLab and FigShare (https://gitlab.com/psc_santos/ACACIAandhttps://figshare.com/projects/ACACIA/66485).

Comparative Effects of High-Tech Visual Scene Displays and Low-Tech Isolated Picture Symbols on Engagement From Students With Multiple Disabilities

2019 ◽  
Vol 50 (4) ◽  
pp. 693-702 ◽  
Author(s):  
Christine Holyfield ◽  
Sydney Brooks ◽  
Allison Schluterman
Keyword(s):  
Language Learning ◽  
Augmentative And Alternative Communication ◽  
Visual Analysis ◽  
Multiple Disabilities ◽  
Visual Scene ◽  
Future Research ◽  
High Tech ◽  
Single Subject ◽  
Wide Range ◽  
The Difference

Purpose Augmentative and alternative communication (AAC) is an intervention approach that can promote communication and language in children with multiple disabilities who are beginning communicators. While a wide range of AAC technologies are available, little is known about the comparative effects of specific technology options. Given that engagement can be low for beginning communicators with multiple disabilities, the current study provides initial information about the comparative effects of 2 AAC technology options—high-tech visual scene displays (VSDs) and low-tech isolated picture symbols—on engagement. Method Three elementary-age beginning communicators with multiple disabilities participated. The study used a single-subject, alternating treatment design with each technology serving as a condition. Participants interacted with their school speech-language pathologists using each of the 2 technologies across 5 sessions in a block randomized order. Results According to visual analysis and nonoverlap of all pairs calculations, all 3 participants demonstrated more engagement with the high-tech VSDs than the low-tech isolated picture symbols as measured by their seconds of gaze toward each technology option. Despite the difference in engagement observed, there was no clear difference across the 2 conditions in engagement toward the communication partner or use of the AAC. Conclusions Clinicians can consider measuring engagement when evaluating AAC technology options for children with multiple disabilities and should consider evaluating high-tech VSDs as 1 technology option for them. Future research must explore the extent to which differences in engagement to particular AAC technologies result in differences in communication and language learning over time as might be expected.

Understanding Limited Use of Amplification in Infants and Children Who Are Hard of Hearing

2015 ◽  
Vol 25 (1) ◽  
pp. 15-23 ◽  
Author(s):  
Ryan W. McCreery ◽  
Elizabeth A. Walker ◽  
Meredith Spratford
Keyword(s):  
Hard Of Hearing ◽  
Hearing Aid ◽  
Management Strategies ◽  
Infants And Children ◽  
Future Research ◽  
Research Needs ◽  
Case Examples ◽  
Hearing Aid Use ◽  
Wide Range ◽  
In Infants And Children

The effectiveness of amplification for infants and children can be mediated by how much the child uses the device. Existing research suggests that establishing hearing aid use can be challenging. A wide range of factors can influence hearing aid use in children, including the child's age, degree of hearing loss, and socioeconomic status. Audiological interventions, including using validated prescriptive approaches and verification, performing on-going training and orientation, and communicating with caregivers about hearing aid use can also increase hearing aid use by infants and children. Case examples are used to highlight the factors that influence hearing aid use. Potential management strategies and future research needs are also discussed.

The Attenuation of Very Low Frequency Brain Oscillations in Transitions from a Rest State to Active Attention

2009 ◽  
Vol 23 (4) ◽  
pp. 191-198 ◽  
Author(s):  
Suzannah K. Helps ◽  
Samantha J. Broyd ◽  
Christopher J. James ◽  
Anke Karl ◽  
Edmund J. S. Sonuga-Barke
Keyword(s):  
Brain Activity ◽  
Sampling Rate ◽  
Low Frequency ◽  
Future Research ◽  
Adhd Symptoms ◽  
Default Mode ◽  
Very Low Frequency ◽  
Wide Range ◽  
Interference Hypothesis ◽  
Mode Interference

Background: The default mode interference hypothesis ( Sonuga-Barke & Castellanos, 2007 ) predicts (1) the attenuation of very low frequency oscillations (VLFO; e.g., .05 Hz) in brain activity within the default mode network during the transition from rest to task, and (2) that failures to attenuate in this way will lead to an increased likelihood of periodic attention lapses that are synchronized to the VLFO pattern. Here, we tested these predictions using DC-EEG recordings within and outside of a previously identified network of electrode locations hypothesized to reflect DMN activity (i.e., S3 network; Helps et al., 2008 ). Method: 24 young adults (mean age 22.3 years; 8 male), sampled to include a wide range of ADHD symptoms, took part in a study of rest to task transitions. Two conditions were compared: 5 min of rest (eyes open) and a 10-min simple 2-choice RT task with a relatively high sampling rate (ISI 1 s). DC-EEG was recorded during both conditions, and the low-frequency spectrum was decomposed and measures of the power within specific bands extracted. Results: Shift from rest to task led to an attenuation of VLFO activity within the S3 network which was inversely associated with ADHD symptoms. RT during task also showed a VLFO signature. During task there was a small but significant degree of synchronization between EEG and RT in the VLFO band. Attenuators showed a lower degree of synchrony than nonattenuators. Discussion: The results provide some initial EEG-based support for the default mode interference hypothesis and suggest that failure to attenuate VLFO in the S3 network is associated with higher synchrony between low-frequency brain activity and RT fluctuations during a simple RT task. Although significant, the effects were small and future research should employ tasks with a higher sampling rate to increase the possibility of extracting robust and stable signals.

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