scholarly journals Advanced Biological Imaging for Intracellular Micromanipulation: Methods and Applications

2020 ◽  
Vol 10 (20) ◽  
pp. 7308
Author(s):  
Wendi Gao ◽  
Libo Zhao ◽  
Zhuangde Jiang ◽  
Dong Sun
Keyword(s):  
Success Rate ◽  
Genome Editing ◽  
Biomedical Research ◽  
Genetic Diagnosis ◽  
Biological Imaging ◽  
Robotic Systems ◽  
Current Development ◽  
Insufficient Information ◽  
Processing Methods ◽  
Future Extension

Intracellular micromanipulation assisted by robotic systems has valuable applications in biomedical research, such as genetic diagnosis and genome-editing tasks. However, current studies suffer from a low success rate and a large operation damage because of insufficient information on the operation information of targeted specimens. The complexity of the intracellular environment causes difficulties in visualizing manipulation tools and specimens. This review summarizes and analyzes the current development of advanced biological imaging sampling and computational processing methods in intracellular micromanipulation applications. It also discusses the related limitations and future extension, providing an important reference about this field.

Recent Progress in Chemosensors Using Aldehyde-bearing Fluorophores for the Detection of Specific Analytes and their Bioimaging

2019 ◽  
Vol 26 (21) ◽  
pp. 4003-4028 ◽  
Author(s):  
Fangjun Huo ◽  
Yaqiong Zhang ◽  
Caixia Yin
Keyword(s):  
Biomedical Research ◽  
Nucleophilic Addition ◽  
Recent Progress ◽  
Biological Imaging ◽  
Tissue Imaging ◽  
Fluorescence Probes ◽  
Biological Applications ◽  
Convenient Procedure ◽  
Nucleophilic Mechanism

In recent years, aldehyde-appended fluorescence probes have attracted increasing attention. Fluorescent biological imaging includes many modern applications for cell and tissue imaging in biomedical research. Meanwhile, the nucleophilic mechanism is a very simple and convenient procedure for the preparation of aldehyde-sensing probes. This tutorial review focuses on aldehyde-bearing chemosensors based on nucleophilic addition mechanism with biological applications.

scholarly journals Genome editing in large animals: current status and future prospects

2019 ◽  
Vol 6 (3) ◽  
pp. 402-420 ◽  
Author(s):  
Jianguo Zhao ◽  
Liangxue Lai ◽  
Weizhi Ji ◽  
Qi Zhou
Keyword(s):  
Regenerative Medicine ◽  
Genome Editing ◽  
Biomedical Research ◽  
Human Disease ◽  
Gene Editing ◽  
Disease Modeling ◽  
Current Status ◽  
Future Prospects ◽  
Recent Advances ◽  
Large Animals

AbstractLarge animals (non-human primates, livestock and dogs) are playing important roles in biomedical research, and large livestock animals serve as important sources of meat and milk. The recently developed programmable DNA nucleases have revolutionized the generation of gene-modified large animals that are used for biological and biomedical research. In this review, we briefly introduce the recent advances in nuclease-meditated gene editing tools, and we outline these editing tools’ applications in human disease modeling, regenerative medicine and agriculture. Additionally, we provide perspectives regarding the challenges and prospects of the new genome editing technology.

scholarly journals Embryo-Based Large Fragment Knock-in in Mammals: Why, How and What’s Next

Genes ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 140 ◽  
Author(s):  
Steven Erwood ◽  
Bin Gu
Keyword(s):  
Animal Model ◽  
Animal Models ◽  
Genome Editing ◽  
Biomedical Research ◽  
Future Development ◽  
Genetic Modification ◽  
Technological Innovations ◽  
Model Generation ◽  
Large Fragment ◽  
Diverse Range

Endonuclease-mediated genome editing technologies, most notably CRISPR/Cas9, have revolutionized animal genetics by allowing for precise genome editing directly through embryo manipulations. As endonuclease-mediated model generation became commonplace, large fragment knock-in remained one of the most challenging types of genetic modification. Due to their unique value in biological and biomedical research, however, a diverse range of technological innovations have been developed to achieve efficient large fragment knock-in in mammalian animal model generation, with a particular focus on mice. Here, we first discuss some examples that illustrate the importance of large fragment knock-in animal models and then detail a subset of the recent technological advancements that have allowed for efficient large fragment knock-in. Finally, we envision the future development of even larger fragment knock-ins performed in even larger animal models, the next step in expanding the potential of large fragment knock-in in animal models.

scholarly journals Microhomologies are prevalent at Cas9-induced larger deletions

2019 ◽  
Vol 47 (14) ◽  
pp. 7402-7417 ◽  
Author(s):  
Dominic D G Owens ◽  
Adam Caulder ◽  
Vincent Frontera ◽  
Joe R Harman ◽  
Alasdair J Allan ◽  
...  
Keyword(s):  
Genome Editing ◽  
Biomedical Research ◽  
High Frequency ◽  
Cultured Cells ◽  
Mouse Embryos ◽  
Deletion Breakpoint ◽  
End Joining ◽  
Crispr System ◽  
Cas9 Nuclease ◽  
Target Sites

Abstract The CRISPR system is widely used in genome editing for biomedical research. Here, using either dual paired Cas9D10A nickases or paired Cas9 nuclease we characterize unintended larger deletions at on-target sites that frequently evade common genotyping practices. We found that unintended larger deletions are prevalent at multiple distinct loci on different chromosomes, in cultured cells and mouse embryos alike. We observed a high frequency of microhomologies at larger deletion breakpoint junctions, suggesting the involvement of microhomology-mediated end joining in their generation. In populations of edited cells, the distribution of larger deletion sizes is dependent on proximity to sgRNAs and cannot be predicted by microhomology sequences alone.

scholarly journals CRISPR-Cas9 in gene therapy: much control on breaking, little control on repairing

2015 ◽  
Author(s):  
Kaveh Daneshvar
Keyword(s):  
Gene Therapy ◽  
Dna Repair ◽  
Immune Responses ◽  
Genome Editing ◽  
Biomedical Research ◽  
Target Cells ◽  
Target Dna ◽  
In Vivo Delivery ◽  
Target Effects

Recent advances in CRISPR-Cas9 genome editing tool have made great promises to basic and biomedical research as well as gene therapy. Efforts to make the CRISPR-Cas9 system applicable in gene therapy are largely focused on two aspects: 1) increasing the specificity of this system by eliminating off-target effects, and 2) optimizing in vivo delivery of the CRISPR-Cas9 DNA constructs to target cells and limiting the expression of Cas9 and gRNA to prevent toxicity immune responses. However, there is an unnoted but crucial consideration about the mode of DNA repair at the lesion caused by CRISPR-Cas9. In this commentary, I briefly highlight recent publications on in vivo use of the CRISPR-Cas9 system in gene therapy. I then discuss the undesired on-target DNA repair events that can occur as a result of the activity of CRISPR-Cas9. Overall, this commentary underscores the need for more study on controlled DNA repair in systems targeted with CRISPR-Cas9 genome editing tools.

scholarly journals Robotics in neurointerventional surgery: a systematic review of the literature

2021 ◽  
pp. neurintsurg-2021-018096
Author(s):  
William Crinnion ◽  
Ben Jackson ◽  
Avnish Sood ◽  
Jeremy Lynch ◽  
Christos Bergeles ◽  
...  
Keyword(s):  
Systematic Review ◽  
Carotid Artery ◽  
Success Rate ◽  
Cerebral Angiography ◽  
Clinical Success ◽  
Robotic Systems ◽  
Occupational Hazards ◽  
Review Of The Literature ◽  
Neurointerventional Procedures ◽  
Manual Assistance

BackgroundRobotically performed neurointerventional surgery has the potential to reduce occupational hazards to staff, perform intervention with greater precision, and could be a viable solution for teleoperated neurointerventional procedures.ObjectiveTo determine the indication, robotic systems used, efficacy, safety, and the degree of manual assistance required for robotically performed neurointervention.MethodsWe conducted a systematic review of the literature up to, and including, articles published on April 12, 2021. Medline, PubMed, Embase, and Cochrane register databases were searched using medical subject heading terms to identify reports of robotically performed neurointervention, including diagnostic cerebral angiography and carotid artery intervention.ResultsA total of 8 articles treating 81 patients were included. Only one case report used a robotic system for intracranial intervention, the remaining indications being cerebral angiography and carotid artery intervention. Only one study performed a comparison of robotic and manual procedures. Across all studies, the technical success rate was 96% and the clinical success rate was 100%. All cases required a degree of manual assistance. No studies had clearly defined patient selection criteria, reference standards, or index tests, preventing meaningful statistical analysis.ConclusionsGiven the clinical success, it is plausible that robotically performed neurointerventional procedures will eventually benefit patients and reduce occupational hazards for staff; however, there is no high-level efficacy and safety evidence to support this assertion. Limitations of current robotic systems and the challenges that must be overcome to realize the potential for remote teleoperated neurointervention require further investigation.

Genome editing of potato using CRISPR technologies: current development and future prospective

2019 ◽  
Vol 139 (2) ◽  
pp. 403-416 ◽  
Author(s):  
Sarbesh Das Dangol ◽  
Abdellah Barakate ◽  
Jennifer Stephens ◽  
Mehmet Emin Çalıskan ◽  
Allah Bakhsh
Keyword(s):  
Genome Editing ◽  
Current Development

scholarly journals Efficient Genome Editing in Bacillus licheniformis Mediated by a Conditional CRISPR/Cas9 System

2020 ◽  
Vol 8 (5) ◽  
pp. 754
Author(s):  
Youran Li ◽  
Hanrong Wang ◽  
Liang Zhang ◽  
Zhongyang Ding ◽  
Sha Xu ◽  
...  
Keyword(s):  
Success Rate ◽  
Genome Editing ◽  
Bacillus Licheniformis ◽  
Genetic Manipulation ◽  
Inducible Promoter ◽  
Bacterial Cells ◽  
Protospacer Adjacent Motif ◽  
Constitutive Overexpression ◽  
Short Palindromic Repeat ◽  
Overexpression System

Bacillus licheniformis is widely used to produce multiple enzymes and chemicals in industrial fermentation. It is also an organism that is hard to genetically manipulate, which is mainly attributed to its extremely low transformation efficiency. The lack of genetic modification technology severely limits its further application. In this study, an all-in-one conditional clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 plasmid was developed for B. licheniformis with the cas9 gene under the control of a xylose-inducible promoter. By means of this design, the expression of the cas9 gene could be repressed without xylose, which significantly improved the transformation ratio from less than 0.1 cfu/μg to 2.42 cfu/μg DNA. Compared with this conditional system, a constitutive overexpression system led to significant growth retardation in bacterial cells. Both the biomass and specific growth rate decreased greatly. After transformation, successful genome editing could be triggered by 0.5% xylose. When the α-amylase gene amyL was used as a genomic target, the efficiencies of its disruption using three different protospacer-adjacent motif (PAM) sequences were 64.3%, 70.9%, and 47.1%, respectively. Moreover, temperature plays a pivotal role in the function of the constructed CRISPR system. The maximum success rate reached 97% at 20 °C, while higher temperatures negatively impacted the function of the system. These results suggested that the design with a cas9 gene under the strict control of a xylose-inducible promoter significantly improved the success rate of genome editing in this host. This work contributes to the development of genetic manipulation and furthers the use of B. licheniformis as an efficient industrial workhorse.

scholarly journals THE JOURNEY OF CRISPR-CAS9 FROM BACTERIAL DEFENSE MECHANISM TO A GENE EDITING TOOL IN BOTH ANIMALS AND PLANTS

2020 ◽  
Vol 2020 (1) ◽  
Author(s):  
T Tahir ◽  
Q Ali ◽  
MS Rashid ◽  
A Malik
Keyword(s):  
Immune System ◽  
Genetic Engineering ◽  
Success Rate ◽  
Genome Editing ◽  
Zinc Finger ◽  
Gene Editing ◽  
Defense Mechanism ◽  
Zinc Finger Nucleases ◽  
Transcription Activator

Today we can use multiple of endonucleases for genome editing which has become very important and used in number of applications. We use sequence specific molecular scissors out of which, most important are mega nucleases, zinc finger nucleases, TALENS (Transcription Activator Like-Effector Nucleases) and CRISPR-Cas9 which is currently the most famous due to a number of reasons, they are cheap, easy to build, very specific in nature and their success rate in plants and animals is also high. Who knew that one day these CRISPR discovered as a part of immune system of bacteria will be this much worthwhile in the field of genetic engineering? This review interprets the science behind their mechanism and how several advancements were made with the passage of time to make them more efficient for the assigned job.

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