Current advances of CRISPR-Cas technology in cell therapy

Qiu, Houyuan; Ji, Rui-Jin; Zhang, Ying

doi:10.1016/j.cellin.2022.100067

Cited by 21 publications

(8 citation statements)

References 184 publications

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“…This treatment is expected to work without sacrificing cell viability or function, allowing for inhibition of a heretofore undruggable intracellular checkpoint cytokine-inducible SH2 containing protein (CISH) [ 4 ]. A clinical trial has also been initiated to treat sickle cell disease using the adenine base editor (ABE) (NCT05456880) [ 165 ].…”

Section: Current Landscape Of Mrna-based Drug Pipelinementioning

confidence: 99%

mRNA-based vaccines and therapeutics: an in-depth survey of current and upcoming clinical applications

Wang,

Kumari,

Chen

et al. 2023

J Biomed Sci

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AbstractmRNA-based drugs have tremendous potential as clinical treatments, however, a major challenge in realizing this drug class will promise to develop methods for safely delivering the bioactive agents with high efficiency and without activating the immune system. With regard to mRNA vaccines, researchers have modified the mRNA structure to enhance its stability and promote systemic tolerance of antigenic presentation in non-inflammatory contexts. Still, delivery of naked modified mRNAs is inefficient and results in low levels of antigen protein production. As such, lipid nanoparticles have been utilized to improve delivery and protect the mRNA cargo from extracellular degradation. This advance was a major milestone in the development of mRNA vaccines and dispelled skepticism about the potential of this technology to yield clinically approved medicines. Following the resounding success of mRNA vaccines for COVID-19, many other mRNA-based drugs have been proposed for the treatment of a variety of diseases. This review begins with a discussion of mRNA modifications and delivery vehicles, as well as the factors that influence administration routes. Then, we summarize the potential applications of mRNA-based drugs and discuss further key points pertaining to preclinical and clinical development of mRNA drugs targeting a wide range of diseases. Finally, we discuss the latest market trends and future applications of mRNA-based drugs.

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Section: Current Landscape Of Mrna-based Drug Pipelinementioning

confidence: 99%

mRNA-based vaccines and therapeutics: an in-depth survey of current and upcoming clinical applications

Wang,

Kumari,

Chen

et al. 2023

J Biomed Sci

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“…Among them, SpCas9 is the most widely used due to its high efficiency in gene editing and simple protospacer adjacent motif (PAM) sequence (5`-NGG-3`) (Fig. 2 A) [ 62 ]. The gRNA for SpCas9 is comprised of a CRISPR RNA (crRNA) with a 20-nucleotide (20-nt) sequence recognizing target DNA sites and a trans-activating CRISPR RNA (tracrRNA), which provides a scaffold for binding to the Cas9 nuclease.…”

Section: Introductionmentioning

confidence: 99%

Gene editing technology to improve antitumor T-cell functions in adoptive immunotherapy

Ito,

Inoue,

Kagoya

2024

Inflamm Regener

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Adoptive immunotherapy, in which tumor-reactive T cells are prepared in vitro for adoptive transfer to the patient, can induce an objective clinical response in specific types of cancer. In particular, chimeric antigen receptor (CAR)-redirected T-cell therapy has shown robust responses in hematologic malignancies. However, its efficacy against most of the other tumors is still insufficient, which remains an unmet medical need. Accumulating evidence suggests that modifying specific genes can enhance antitumor T-cell properties. Epigenetic factors have been particularly implicated in the remodeling of T-cell functions, including changes to dysfunctional states such as terminal differentiation and exhaustion. Genetic ablation of key epigenetic molecules prevents the dysfunctional reprogramming of T cells and preserves their functional properties.Clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)-based gene editing is a valuable tool to enable efficient and specific gene editing in cultured T cells. A number of studies have already identified promising targets to improve the therapeutic efficacy of CAR-T cells using genome-wide or focused CRISPR screening. In this review, we will present recent representative findings on molecular insights into T-cell dysfunction and how genetic modification contributes to overcoming it. We will also discuss several technical advances to achieve efficient gene modification using the CRISPR and other novel platforms.

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“…The clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein (Cas) system allows for a wide range of applications for gene modification when guided by RNA and in the presence of protospacer-adjacent motif (PAM) sequence [1][2][3][4][5][6][7][8]. However, crRNA-DNA mismatches may also induce cleavage activity of Cas nucleases, resulting in undesired cleavage at off-target sites [9][10][11][12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Engineering of Cas12a nuclease variants with enhanced genome-editing specificity

Chen,

Zhou,

Liu

et al. 2024

PLoS Biol

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The clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a system is a powerful tool in gene editing; however, crRNA-DNA mismatches might induce unwanted cleavage events, especially at the distal end of the PAM. To minimize this limitation, we engineered a hyper fidelity AsCas12a variant carrying the mutations S186A/R301A/T315A/Q1014A/K414A (termed HyperFi-As) by modifying amino acid residues interacting with the target DNA and crRNA strand. HyperFi-As retains on-target activities comparable to wild-type AsCas12a (AsCas12aWT) in human cells. We demonstrated that HyperFi-As has dramatically reduced off-target effects in human cells, and HyperFi-As possessed notably a lower tolerance to mismatch at the position of the PAM-distal region compared with the wild type. Further, a modified single-molecule DNA unzipping assay at proper constant force was applied to evaluate the stability and transient stages of the CRISPR/Cas ribonucleoprotein (RNP) complex. Multiple states were sensitively detected during the disassembly of the DNA-Cas12a-crRNA complexes. On off-target DNA substrates, the HyperFi-As-crRNA was harder to maintain the R-loop complex state compared to the AsCas12aWT, which could explain exactly why the HyperFi-As has low off-targeting effects in human cells. Our findings provide a novel version of AsCas12a variant with low off-target effects, especially capable of dealing with the high off-targeting in the distal region from the PAM. An insight into how the AsCas12a variant behaves at off-target sites was also revealed at the single-molecule level and the unzipping assay to evaluate multiple states of CRISPR/Cas RNP complexes might be greatly helpful for a deep understanding of how CRISPR/Cas behaves and how to engineer it in future.

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Current advances of CRISPR-Cas technology in cell therapy

Cited by 21 publications

References 184 publications

mRNA-based vaccines and therapeutics: an in-depth survey of current and upcoming clinical applications

mRNA-based vaccines and therapeutics: an in-depth survey of current and upcoming clinical applications

Gene editing technology to improve antitumor T-cell functions in adoptive immunotherapy

Engineering of Cas12a nuclease variants with enhanced genome-editing specificity

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