Challenges of CRISPR-based Gene Editing in Primary T Cells

Rezalotfi, Alaleh; Fritz, Lea; Förster, Reinhold; Bošnjak, Berislav

doi:10.20944/preprints202201.0137.v1

Cited by 4 publications

(3 citation statements)

References 156 publications

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“…According to the research on inhibitory receptors and immune checkpoint such as PD1, TGF beta and CTLA-4 important hallmarks of those in cancer treatment were found out. Inhibitory receptors (iRs) are important in function of adaptive immune cells and in T cell exhaustion which is a common event in cancer [38,39] by blocking checkpoint molecules re-establishing the potency of T cells is one important part in the cancer treatment process via immune-checkpoint therapy [40,41]. CRISPR is a good safeguarding tool to derange immune checkpoint by protecting from checkpoint inhibition via knock out immune checkpoint molecules in CAR T cells.…”

Section: How Crispr Is Used In Car T Cell Therapymentioning

confidence: 99%

Molecular and therapeutic effect of CRISPR in treating cancer

2023

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Cancer has become one of the common causes of mortality around the globe due to mutations in the genome which allows rapid growth of cells uncontrollably without repairing DNA errors. Cancers could arise due alterations in DNA repair mechanisms (errors in mismatch repair genes), activation of oncogenes and inactivation of tumor suppressor genes. Each cancer type is different and each individual has a unique genetic change which leads them to cancer. Studying genetic and epigenetic alterations in the genome leads to understanding the underlying features. CAR T therapy over other immunotherapies such as monoclonal antibodies, immune checkpoint inhibitors, cancer vaccines and adoptive cell therapies has been widely used to treat cancer in recent days and gene editing has now become one of the promising treatments for many genetic diseases. This tool allows scientists to change the genome by adding, removing or altering genetic material of an organism. Due to advance in genetics and novel molecular techniques such as CRISPR, TALEN these genes can be edited in such a way that their original function could be replaced which in turn improved the treatment possibilities and can be used against malignancies and even cure cancer in future along with CAR T cell therapy due to the specific recognition and attacking of tumor.

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Section: How Crispr Is Used In Car T Cell Therapymentioning

confidence: 99%

Molecular and therapeutic effect of CRISPR in treating cancer

2023

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“…In conclusion, the overall efficiency of knock-in seems to remain lower than the knockout efficiency, using CRISPR-Cas9. Moreover, delivery of CRISPR-Cas9 through viral vectors requires in vitro T cell activation and culture [41,42]. However, the use of electroporation (EP) methods to deliver Cas9-gRNA protein complex in knockout studies demonstrated the potential to overcome this issue and to achieve gene editing without in vitro T cell activation [42,43].…”

Section: Talens Crispr/casmentioning

confidence: 99%

Current strategies employed in the manipulation of gene expression for clinical purposes

et al. 2022

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Abnormal gene expression level or expression of genes containing deleterious mutations are two of the main determinants which lead to genetic disease. To obtain a therapeutic effect and thus to cure genetic diseases, it is crucial to regulate the host’s gene expression and restore it to physiological conditions. With this purpose, several molecular tools have been developed and are currently tested in clinical trials. Genome editing nucleases are a class of molecular tools routinely used in laboratories to rewire host’s gene expression. Genome editing nucleases include different categories of enzymes: meganucleses (MNs), zinc finger nucleases (ZFNs), clustered regularly interspaced short palindromic repeats (CRISPR)- CRISPR associated protein (Cas) and transcription activator-like effector nuclease (TALENs). Transposable elements are also a category of molecular tools which includes different members, for example Sleeping Beauty (SB), PiggyBac (PB), Tol2 and TcBuster. Transposons have been used for genetic studies and can serve as gene delivery tools. Molecular tools to rewire host’s gene expression also include episomes, which are divided into different categories depending on their molecular structure. Finally, RNA interference is commonly used to regulate gene expression through the administration of small interfering RNA (siRNA), short hairpin RNA (shRNA) and bi-functional shRNA molecules. In this review, we will describe the different molecular tools that can be used to regulate gene expression and discuss their potential for clinical applications. These molecular tools are delivered into the host's cells in the form of DNA, RNA or protein using vectors that can be grouped into physical or biochemical categories. In this review we will also illustrate the different types of payloads that can be used, and we will discuss recent developments in viral and non-viral vector technology.

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“…For autologous CAR-T, this incorporates trouble getting expansive numbers of T cells from patients that are lymphopenic due to other medicines and long timeframes for creating an adequate restorative measurements [65].For allogeneic CAR-T, the solid donor-derived cells can be rejected by the understanding resistant framework, cause harmfulness, or initiate GvHD [66]. Shockingly, a few cancers are able to maintain a strategic distance from pulverization by CAR-T cells by overexpressing certain proteins, like modified cell passing protein 1 (PD-1), on their surface [67] .CRISPR-Cas9 quality altering has expanded the security and viability of CAR-T cell therapies in an assortment of ways [68] .Firstly, CRISPR can be utilized to thump within the CAR in a focused on way -for illustration, to a safe harbor location within the genome -to guarantee adequate, long-term expression of the receptor on the cell surface [69] .Furthermore, CRISPR can thump out certain qualities in CAR-T cells to extend their cancerkilling movement .Thirdly, it can be utilized to form alters that minimize CAR-T cell fatigue and increment their long-term multiplication [70].At long last, CRISPR alters can be utilized to produce all inclusive, 'off-the-shelf' CAR-T cells from actuated pluripotent stem cells (iPSCs), invalidating collecting confinements, long hold up times, and harmfulness issues [71] .…”

Section: Til and Tcr Cell Therapiesmentioning

confidence: 99%

CRISPR and CAR-T, NK Current application and future perspective

khoshandam¹,

Khoshandam

Soltaninejad

et al. 2022

Preprint

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Chimeric Antigen Receptor T-cells represent a breakthrough in personalized cancer therapy. In this strategy, synthetic receptors comprised of antigen recognition, signaling, and stimulatory domains are used to reprogram T-cells to target tumor cells for destruction. Despite the success of this approach in refractory B-cell malignancies, optimal potency of CAR T-cell therapy for many other cancers, particularly solid tumors, has not been achieved. NK cells are powerful cytotoxic lymphocytes specialized in recognizing and dispensing with changed cells, and in coordinating versatile anti-tumor immunity.NK cells are as a rule practically depleted within the tumor microenvironment. In like manner, current investigate endeavors center on exactness designing of CAR T-cells with routine CRISPR-Cas9 frameworks or novel editors that can introduce craved hereditary changes with or without presentation of a double-stranded break into the genome. These instruments and methodologies can be specifically connected to focusing on negative controllers of T-cell work, coordinating helpful transgenes to particular genomic loci, and producing reproducibly secure and powerful allogeneic widespread CAR T-cell items for on-demand cancer immunotherapy. The revelation and improvement of the CRISPR/Cas9 innovation offer an adaptable and proficient gene-editing capability in tweaking different pathways that intercede NK cell fatigue and in outfitting NK cells with novel chimeric antigen receptors to particularly target tumor cells. Despite the tall productivity in its geneediting capability, trouble within the conveyance of the CRISPR/Cas9 framework remains a major bottleneck for its restorative applications, especially for NK cells.This review assesses a few of the progressing and future bearings of combining next-generation CRISPR-Cas9 quality altering with manufactured science to optimize CAR T-cell and NK cell treatment for future clinical trials toward the foundation of a modern cancer treatment parade.

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Challenges of CRISPR-based Gene Editing in Primary T Cells

Cited by 4 publications

References 156 publications

Molecular and therapeutic effect of CRISPR in treating cancer

Molecular and therapeutic effect of CRISPR in treating cancer

Current strategies employed in the manipulation of gene expression for clinical purposes

CRISPR and CAR-T, NK Current application and future perspective

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