Applications of CRISPR/Cas gene-editing technology in yeast and fungi

2021 ◽  
Vol 204 (1) ◽  
Author(s):  
Binyou Liao ◽  
Xi Chen ◽  
Xuedong Zhou ◽  
Yujie Zhou ◽  
Yangyang Shi ◽  
...  
Keyword(s):  
Gene Editing ◽  
Cas Gene

scholarly journals The Diversity of Genetic Outcomes from CRISPR/Cas Gene Editing is Regulated by the Length of the Symmetrical Donor DNA Template

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1160
Author(s):  
Amanda M. Hewes ◽  
Brett M. Sansbury ◽  
Eric B. Kmiec
Keyword(s):  
Gene Editing ◽  
Bacterial Cells ◽  
Knock Out ◽  
Viral Genomes ◽  
Homology Directed Repair ◽  
Established Cell ◽  
Dna Template ◽  
Donor Template ◽  
Eukaryotic Genomes ◽  
Cas Gene

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas gene editing systems have enabled molecular geneticists to manipulate prokaryotic and eukaryotic genomes with greater efficiency and precision. CRISPR/Cas provides adaptive immunity in bacterial cells by degrading invading viral genomes. By democratizing this activity into human cells, it is possible to knock out specific genes to disable their function and repair errors. The latter of these activities requires the participation of a single-stranded donor DNA template that provides the genetic information to execute correction in a process referred to as homology directed repair (HDR). Here, we utilized an established cell-free extract system to determine the influence that the donor DNA template length has on the diversity of products from CRISPR-directed gene editing. This model system enables us to view all outcomes of this reaction and reveals that donor template length can influence the efficiency of the reaction and the categories of error-prone products that accompany it. A careful measurement of the products revealed a category of error-prone events that contained the corrected template along with insertions and deletions (indels). Our data provides foundational information for those whose aim is to translate CRISPR/Cas from bench to bedside.

scholarly journals The technical risks of human gene editing

2019 ◽  
Vol 34 (11) ◽  
pp. 2104-2111 ◽  
Author(s):  
Benjamin Davies
Keyword(s):  
Genome Editing ◽  
Gene Editing ◽  
Human Gene ◽  
Ethical Concerns ◽  
Technical Risks ◽  
Genomic Mutation ◽  
Cas Gene

Abstract A recent report from Dr He Jiankui concerning the birth of twin girls harbouring mutations engineered by CRISPR/Cas nucleases has been met with international condemnation. Beside the serious ethical concerns, there are known technical risks associated with CRISPR/Cas gene editing which further raise questions about how these events could have been allowed to occur. Numerous studies have reported unexpected genomic mutation and mosaicism following the use of CRISPR/Cas nucleases, and it is currently unclear how prevalent these disadvantageous events are and how robust and sensitive the strategies to detect these unwanted events may be. Although Dr Jiankui’s study appears to have involved certain checks to ascertain these risks, the decision to implant the manipulated embryos, given these unknowns, must nonetheless be considered reckless. Here I review the technical concerns surrounding genome editing and consider the available data from Dr Jiankui in this context. Although the data remains unpublished, preventing a thorough assessment of what was performed, it seems clear that the rationale behind the undertaking was seriously flawed; the procedures involved substantial technical risks which, when added to the serious ethical concerns, fully justify the widespread criticism that the events have received.

scholarly journals Applications of CRISPR/Cas Gene-Editing Technology in Fungi

2021 ◽  
Author(s):  
Binyou Liao ◽  
Lei Cheng ◽  
Yujie Zhou ◽  
Yangyang Shi ◽  
Xingchen Ye ◽  
...  
Keyword(s):  
Gene Editing ◽  
Yeast Genome ◽  
Transcription Activator ◽  
Single Nucleotide ◽  
Complex Design ◽  
Infection Treatment ◽  
New Gene ◽  
Low Efficiency ◽  
Genomic Editing ◽  
Cas Gene

Abstract Genome editing technology develop fast in recent years. The traditional gene-editing methods, including homologous recombination, zinc finger endonuclease, and transcription activator-like effector nuclease and so on, which have greatly promoted the research of genetics and molecular biology, have gradually showed their limitations such as low efficiency, high error rate, and complex design. In 2012,a new gene-editing technology, the CRISPR/Cas9 system, was setup based on the research of the immune responses to viruses from archaea and bacteria. Due to its advantages of high target efficiency, simple primer design, and wide application, CRISPR/Cas9 system, whose developers are awared the Nobel Prize in Chemistry this year, has become the dominant genomic editing technology in global academia and some pharmaceuticals. Here we briefly introduce the CRISPR/Cas system and its main applications in yeast, filamentous fungi and macrofungi, including single nucleotide, polygene and polyploid editing, yeast chromosome construction, yeast genome and yeast library construction, CRISPRa/CRISPRi-mediated, CRISPR platform of non-traditional yeast and regulation of metabolic pathway, to highlight the possible applications on fungal infection treatment and to promote the transformation and application of the CRISPR/Cas system in fungi.

scholarly journals CRISPR/Cas-based Diagnostics and Gene Therapy

2021 ◽  
Author(s):  
Qiu Meiyu ◽  
Li Pei
Keyword(s):  
Gene Therapy ◽  
Gene Editing ◽  
Ex Vivo ◽  
Point Of Care ◽  
Cost Effective ◽  
Blood Disorders ◽  
Initial Stage ◽  
Cas Gene ◽  
Cas Proteins

Clustered regularly interspaced short palindromic repeats (CRISPR) technology, an easy, rapid, cost-effective, and precise gene-editing technique, has revolutionized diagnostics and gene therapy. Fast and accurate diagnosis of diseases is essential for point-of-care-testing (POCT) and specialized medical institutes. The CRISPR-associated (Cas) proteins system shed light on the new diagnostics methods at point-of-care (POC) owning to its advantages. In addition, CRISPR/Cas-based gene-editing technology has led to various breakthroughs in gene therapy. It has been employed in clinical trials for a variety of untreatable diseases, including cancer, blood disorders, and other syndromes. Currently, the clinical application of CRISPR/Cas has been mainly focused on ex vivo therapies. Recently, tremendous efforts have been made in the development of ex vivo gene therapy based on CRISPR-Cas9. Despite these efforts, in vivo CRISPR/Cas gene therapy is only in its initial stage. Here, we review the milestones of CRISPR/Cas technologies that advanced the field of diagnostics and gene therapy. We also highlight the recent advances of diagnostics and gene therapy based on CRISPR/Cas technology. In the last section, we discuss the strength and significant challenges of the CRISPR/Cas technology for its future clinical usage in diagnosis and gene therapy.

EMT signaling: potential contribution of CRISPR/Cas gene editing

2020 ◽  
Vol 77 (14) ◽  
pp. 2701-2722 ◽  
Author(s):  
Reza Mohammadinejad ◽  
Alessio Biagioni ◽  
Ganesan Arunkumar ◽  
Rebecca Shapiro ◽  
Kun-Che Chang ◽  
...  
Keyword(s):  
Gene Editing ◽  
Potential Contribution ◽  
Cas Gene

The Potential of CRISPR/Cas Gene Editing to Correct Male Infertility

2020 ◽  
pp. 347-367
Author(s):  
Douglas T. Carrell ◽  
Jingtao Guo ◽  
Kenneth I. Aston
Keyword(s):  
Male Infertility ◽  
Gene Editing ◽  
Cas Gene

scholarly journals Engineered CRISPR/Cas13d Sensing hTERT Selectively Inhibits the Progression of Bladder Cancer In Vitro

2021 ◽  
Vol 8 ◽  
Author(s):  
Chengle Zhuang ◽  
Changshui Zhuang ◽  
Qun Zhou ◽  
Xueting Huang ◽  
Yaoting Gui ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Gene Editing ◽  
New Device ◽  
T24 Cells ◽  
Bladder Cancer Cells ◽  
Cas Gene

Aptazyme and CRISPR/Cas gene editing system were widely used for regulating gene expression in various diseases, including cancer. This work aimed to reconstruct CRISPR/Cas13d tool for sensing hTERT exclusively based on the new device OFF-switch hTERT aptazyme that was inserted into the 3’ UTR of the Cas13d. In bladder cancer cells, hTERT ligand bound to aptamer in OFF-switch hTERT aptazyme to inhibit the degradation of Cas13d. Results showed that engineered CRISPR/Cas13d sensing hTERT suppressed cell proliferation, migration, invasion and induced cell apoptosis in bladder cancer 5637 and T24 cells without affecting normal HFF cells. In short, we constructed engineered CRISPR/Cas13d sensing hTERT selectively inhibited the progression of bladder cancer cells significantly. It may serve as a promising specifically effective therapy for bladder cancer cells.

Sign in / Sign up

Export Citation Format

Share Document