A CRISPR view of hematopoietic stem cells: Moving innovative bioengineering into the clinic

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas genome engineering has emerged as a powerful tool to modify precise genomic sequences with unparalleled accuracy and efficiency. Major advances in CRISPR technologies over the last 5 years have fueled the development of novel techniques in hematopoiesis research to interrogate the complexities of hematopoietic stem cell (HSC) biology. In particular, high throughput CRISPR based screens using various “flavors” of Cas coupled with sequencing and/or functional outputs are becoming increasingly efficient and accessible. In this review, we discuss recent achievements in CRISPR‐mediated genomic engineering and how these new tools have advanced the understanding of HSC heterogeneity and function throughout life. Additionally, we highlight how these techniques can be used to answer previously inaccessible questions and the challenges to implement them. Finally, we focus on their translational potential to both model and treat hematological diseases in the clinic.

Promising Stem Cell therapy in the Management of HIV and AIDS: A Narrative Review

“…High-throughput CRISPR screenings using many “new flavours” of Cas and sequential and/or functional outcomes, in specific, have become more effective and practical. 104 , 105 …”

“…Non-homologous end joining can cause insertion–deletion (indel) substitutions at the target location, whereas homology-directed repair can use a template DNA to insert new genetic material. 104 , 106 …”

“…Whilst single-gene CRISPR/Cas9 programming is a significant tool for testing gene function in primary hematopoietic cells, high-throughput screenings potentially offer CRISPR/Cas9 an even greater advantage in hematopoietic research. 104 …”

“…CRISPR has also been used to target CCR5 in CD34+ HSPCs in an effort to make immune cells resistant to HIV infection, as CCR5 is an important coreceptor for HIV infection. 104 …”

“…In the future, CRISPR/SaCas9 and Ribonucleoprotein (RNP) administration should be explored in the further investigations. 108 As well as some different title studies were explained the effectiveness of the CRISPR gene editing technology on the management of HIV/AIDS including: CRISPR view of hematopoietic stem cells: Moving innovative bioengineering into the clinic, 104 CRISPR-Edited Stem Cells in a Patient with HIV and Acute Lymphocytic Leukaemia, 109 Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice, 110 Extinction of all infectious HIV in cell culture by the CRISPR-Cas12a system with only a single crRNA, 111 HIV-specific humoral immune responses by CRISPR/Cas9-edited B cells, 112 CRISPR-Cas9 Mediated Exonic Disruption for HIV-1 Elimination, 113 RNA-directed gene editing specifically eradicates latent and prevents new HIV-1 infection, 114 CRISPR/Cas9 Ablation of Integrated HIV-1 Accumulates Pro viral DNA Circles with Reformed Long Terminal Repeats, 115 CRISPR-Cas9-mediated gene disruption of HIV-1 co-receptors confers broad resistance to infection in human T cells and humanized mice, 116 Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9, 117 Transient CRISPR-Cas Treatment Can Prevent Reactivation of HIV-1 Replication in a Latently Infected T-Cell Line, 118 CCR5 Gene Disruption via Lentiviral Vectors Expressing Cas9 and Single Guided RNA Renders Cells Resistant to HIV-1 Infection, 119 CRISPR/Cas9-Mediated CCR5 Ablation in Human Hematopoietic Stem/Progenitor Cells Confers HIV-1 Resistance In Vivo. 109 …”