CRISPR/Cascade 9-Mediated Genome Editing-Challenges and Opportunities

Abstract

Clustered Regularly Interspaced Palindromic Repeats (CRISPR) and Cascade 9 (also known as Cas9, CRISPR associated protein 9) confer protection against invading viruses or plasmids. The CRISPR/Cascade 9 system constitutes one of the most powerful genome technologies available to researchers today. So far, this technology has enabled efficient genome editing and modification in several model organisms and has successfully been used in biomedicine and biomedical engineering. However, challenges for efficient and safe genetic manipulation in several organisms persist. Here, we review functional approaches and future challenges associated with the use of the CRISPR/Cascade 9 genome editing system and discuss opportunities, ethical issues and future directions within this field.

Nuclear compartmentalization, dynamics, and function of regulatory DNA sequences

“…Instead, a combination of upcoming approaches for single cell analyses (for review, see Reference ) and biophysical approaches together with microscopic tools, including fluorescence cross‐correlation spectroscopy, will most likely provide successful solutions to analyze the true 3D geometry, compaction, accessibility, and dynamic spatial positioning of active and inactive chromatin structures, including active and inactive TREs. Methods for the selective proteolysis of proteins, as well as for the targeted deletion of genes involved in envisaged molecular mechanisms provide possibilities for functional tests.…”

Recent advancements in exon-skipping therapies using antisense oligonucleotides and genome editing for the treatment of various muscular dystrophies

“…CRISPR system shows a great flexibility in genetic modification but in vivo delivery of CRISPR-Cas9 and non-specific off-target effects remain challenging (Ref. 87). There are many ways CRISPR can be used to treat genetic diseases including homology-directed repair (HDR)-mediated gene correction, single base pair correction, exon deletions and frame correction via insertion and deletion (INDEL) mutations (Refs 22, 23, 88).…”

The renaissance of yeasts as microbial factories in the modern age of biomanufacturing

“…To date, generating these deletion collections was a laborious process limiting their applications to a handful of strains, but several methods have been developed for genome‐scale engineering in a variety of species (e.g., Si et al, ). CRISPR‐Cas9 gene editing has been combined with genetic barcoding to enable efficient, traceable, markerless, and genome‐scale editing (Bao et al, ; Guo et al, ; Roy et al, ). For example, Bao et al () were able to use their CRISPR‐Cas‐based method combined with barcoding to identify a yeast with a 20‐fold increase the tolerance to lactic acid.…”