Tag: CRISPR-Cas9

Transformation of bacterial cells with a plasmid encoding CRISPR-Cas9 machinery is still a necessary step for implementing the technique

May 24, 2021

Since its promulgation in 2012, CRISPR-Cas9 has emerged as the dominant technique for implementing genome editing in a variety of species and cell type. But, how is the technique implemented? What does the technique rely on? In essence, CRISPR-Cas9 genome editing requires a plasmid to encode both the guide RNA and Cas9 protein. This plasmid … More Transformation of bacterial cells with a plasmid encoding CRISPR-Cas9 machinery is still a necessary step for implementing the technique

CRISPR-Cas9 mediated promoter knock-in activates silent biosynthetic gene clusters

April 2, 2021

Even in bacterial species categorized as prolific producers of secondary metabolites, many biosynthetic gene clusters remain silent under standard laboratory cultivation conditions. Hence, to help improve the types of secondary metabolites that could be produced, activation of silent biosynthetic gene clusters has emerged as an important challenge in biotechnology. To this end, many approaches have … More CRISPR-Cas9 mediated promoter knock-in activates silent biosynthetic gene clusters

Profiling cryptic metabolites from microbes

November 27, 2020

Natural products are a rich source of metabolites useful for medicinal uses as antibiotics, anti-inflammatory drugs or anti-cancer agents. Typically obtained as secondary metabolites from microorganisms such as Streptomyces and actinomycetes, these natural products constitute about 35% of all approved medicines by the Food and Drug Administration of U.S. and the European Medicine Agency. However, … More Profiling cryptic metabolites from microbes

CRISPR-Cas9 genome editing relies on transforming cells with a plasmid to effect gene editing

September 25, 2020

Amongst genome editing methods, CRISPR-Cas9 has emerged as the most popular given its ease of use and design and high fidelity of gene editing. Specifically, the key part of the genome editing approach concerns the design of the single stranded guide RNA that help locate the Cas9 endonuclease to a specific region of the genome … More CRISPR-Cas9 genome editing relies on transforming cells with a plasmid to effect gene editing

Anti-CRISPR proteins provide immunity to invading bacteriophages

August 12, 2020

Although commonly known as a genome editing tool, the original function of CRISPR (clustered regularly interspersed short palindromic repeats) is to provide adaptive immunity to bacteria. To do this, CRISPR catalogues short snippets of genetic material from invading bacteriophages into a repeat sequence, which upon transcription, would provide an ensemble of short RNA sequences in … More Anti-CRISPR proteins provide immunity to invading bacteriophages

CRISPR-Cas9 enables systematic investigation of gene function in organisms

September 21, 2019

Originally an adaptive immune system in bacteria, CRISPR-Cas9 (clustered regularly interspersed palindromic repeats – Cas associated protein) has been widely utilized in genome editing applications in a variety of organisms spanning eukaryotes and bacteria. Able to mediate a specific and targeted double strand break to any location in a genome after guidance from a user-designed … More CRISPR-Cas9 enables systematic investigation of gene function in organisms

Cas9 protein may not be able to access DNA encased in heterochromatin

August 2, 2019

CRISPR-Cas9 is the dominant gene editing technology in use. Utilizing the ability of guide RNA to specifically target any stretch of DNA, the system is able to guide the endonuclease Cas9 protein to the location indicated by the guide RNA to generate a double strand break that could serve as a locus for new DNA … More Cas9 protein may not be able to access DNA encased in heterochromatin

Re-domestication of food crops using CRISPR-Cas9 genome editing

May 19, 2019

Selective breeding has played a major role in the evolution of domesticated food crops such as tomatoes. For example, progressive selective breeding by farmers selected for traits useful for garnering a higher price for the food crop as well as enabling ease of harvest. However, significant flavour and nutritional content were lost from the food … More Re-domestication of food crops using CRISPR-Cas9 genome editing

Utility of CRISPR-Cas9 in genome editing lies in the error rate of DNA repair

December 16, 2017

Originally used in defending bacterial cells from attacks by viruses, clustered regularly interspersed short palindromic repeats (CRISPR) has been modified and utilized in editing single bases in DNA in a process known as genome editing. However, efficiency of CRISPR and CRISPR associated protein 9 (Cas9) in executing precise single nucleotide genome editing remains poor due … More Utility of CRISPR-Cas9 in genome editing lies in the error rate of DNA repair