Advances in CRISPR therapeutics

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR) renaissance was catalysed by the discovery that RNA-guided prokaryotic CRISPR-associated (Cas) proteins can create targeted double-strand breaks in mammalian genomes. This finding led to the development of CRISPR systems that harness natural DNA repair mechanisms to repair deficient genes more easily and precisely than ever before. CRISPR has been used to knock out harmful mutant genes and to fix errors in coding sequences to rescue disease phenotypes in preclinical studies and in several clinical trials. However, most genetic disorders result from combinations of mutations, deletions and duplications in the coding and non-coding regions of the genome and therefore require sophisticated genome engineering strategies beyond simple gene knockout. To overcome this limitation, the toolbox of natural and engineered CRISPR–Cas systems has been dramatically expanded to include diverse tools that function in human cells for precise genome editing and epigenome engineering. The application of CRISPR technology to edit the non-coding genome, modulate gene regulation, make precise genetic changes and target infectious diseases has the potential to lead to curative therapies for many previously untreatable diseases.

Myocardial IGF2R is a critical mediator of inflammation and fibrosis after ischemia-reperfusion injury

“…reversibility, and presents a promising approach for treating a diverse range of diseases. 45,46 IGF2R expression was significantly eliminated by CRISPRi which was found to attenuate cell death under hypoxic conditions, yet the molecular pathways involved in this protective effect remain to be determined. Future studies using I/R animal models will be also needed to determine the potential therapeutic effect of this approach.…”

Exploring microbial functional biodiversity at the protein family level—From metagenomic sequence reads to annotated protein clusters

“…The procedures followed are presented from the scope of analyzing assembled contigs; however, the pipelines also support the annotation of amplicons, fragments, and, in the case of MGnify, unassembled reads, by using most of the same tools. Some specific details differentiate among the workflows, as each may use different tools for the same type of annotation, or perform additional analyses; for example, the DOE JGI pipeline also searches for CRISPR elements ( Anzalone et al, 2020 ; Makarova et al, 2020 ; Nidhi et al, 2021 ; Chavez et al, 2022 ; Katti et al, 2022 ; Wang et al, 2022 ) with CRT-CLI ( Bland et al, 2007 ; Clum et al, 2021 ). However, all three workflows follow, more or less, the same procedure, which consists of the following stages: i) the detection of non-coding RNA (ncRNA) genes, ii) the prediction of protein-coding genes, and iii) functional annotation of proteins and taxonomic assignment.…”

X‐ray‐Guided In Situ Genetic Engineering of Macrophages for Sustained Cancer Immunotherapy

“…Due to the advantages of high specificity, efficiency, and simplicity, the CRISPR/Cas9 genome editing system has recently been recognized as an ideal genome editing tool in biomedical research. [40][41][42][43][44][45][46][47][48][49][50][51][52][53] Reprogramming the genome of macrophages using CRISPR/Cas9 has been explored as a strategy, which uses the guide of sgRNA and Cas9 endonuclease to effectively disrupt target genes. Targeting delivery of CRISPR/Cas9 system to macrophages is a promising approach to repolarize TAMs from the protumorigenic M2 phenotype to the antitumor M1 without reconversion and to maintain macrophages in an activated M1 state.…”