Random Insertional Mutagenesis of a Serotype 2 Dengue Virus Clone
Authors: Jeffrey W. Perry and
Andrew W. Tai,
date: 08/20/2018,
view: 1477,
Q&A: 0
[Abstract] Protein tagging is a powerful method of investigating protein function. However, modifying positive-strand RNA virus proteins in the context of viral infection can be particularly difficult as their compact genomes and multifunctional proteins mean even small changes can inactivate or attenuate the virus. Although targeted approaches to ...
CRISPR/Cas Gene Editing of a Large DNA Virus: African Swine Fever Virus
[Abstract] Gene editing of large DNA viruses, such as African swine fever virus (ASFV), has traditionally relied on homologous recombination of a donor plasmid consisting of a reporter cassette with surrounding homologous viral DNA. However, this homologous recombination resulting in the desired modified virus is a rare event. We recently reported the use of ...
Method for CRISPR/Cas9 Mutagenesis in Candida albicans
Authors: Neta Dean and
Henry Ng,
date: 04/20/2018,
view: 4248,
Q&A: 0
[Abstract] Candida albicans is the most prevalent and important human fungal pathogen. The advent of CRISPR as a means of gene editing has greatly facilitated genetic analysis in C. albicans. Here, we describe a detailed step-by-step procedure to construct and analyze C. albicans deletion mutants. This protocol uses plasmids that ...
Single-step Precision Genome Editing in Yeast Using CRISPR-Cas9
[Abstract] Genome modification in budding yeast has been extremely successful largely due to its highly efficient homology-directed DNA repair machinery. Several methods for modifying the yeast genome have previously been described, many of them involving at least two-steps: insertion of a selectable marker and substitution of that marker for the intended ...
Microbial Mutagenicity Assay: Ames Test
[Abstract] The Microbial mutagenicity Ames test is a bacterial bioassay accomplished in vitro to evaluate the mutagenicity of various environmental carcinogens and toxins. While Ames test is used to identify the revert mutations which are present in strains, it can also be used to detect the mutagenicity of environmental samples such as drugs, dyes, ...
Multiple Stepwise Gene Knockout Using CRISPR/Cas9 in Escherichia coli
[Abstract] With the recent implementation of the CRISPR/Cas9 technology as a standard tool for genome editing, laboratories all over the world are undergoing one of the biggest advancements in molecular biology since PCR. The key advantage of this method is its simplicity and universal applicability for species of any phylum. Of particular interest is the ...
Genome Editing in Diatoms Using CRISPR-Cas to Induce Precise Bi-allelic Deletions
[Abstract] Genome editing in diatoms has recently been established for the model species Phaeodactylum tricornutum and Thalassiosira pseudonana. The present protocol, although developed for T. pseudonana, can be modified to edit any diatom genome as we utilize the flexible, modular Golden Gate cloning system. The main steps include ...