Harnessing <scp>high‐throughput</scp> flow cytometric protein and <scp>RNA</scp> detection to enhance <scp>TIL</scp> therapy

Heeren, Julian J Freen-van

doi:10.1002/cyto.a.24323

Cited by 2 publications

(2 citation statements)

References 15 publications

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“…By coupling CRISPR/Casmediated genome editing to other tools, such as Flow-FISH, novel insights have been gained into basic cellular processes, the way infections like SARS-CoV-2 rewire cellular metabolism, and the complex network of interactions of cis-regulatory elements and their target genes. Other potential applications could include screening for CRISPR repair efficiency by making use of template-specific probes, or assessing the effector function of tumor infiltrating lymphocyte products with Flow-FISH [17] after CRISPR-mediated genome editing [18]. However, while using e.g.…”

Section: Authors Used a Pooled Crispr Interference Approach To Identifymentioning

confidence: 99%

Combining CRISPR with Flow‐FISH to study CRISPR‐mediated genome perturbation

Freen‐van Heeren

2023

Cytometry Pt A

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Since the advent of the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) system as a genome editing tool, the ease of studying gene function and the impact thereof on cellular function has increased incrementally. Not surprisingly, the original describers of the CRISPR/Cas system received the 2020 Nobel Prize in Chemistry. Compared to conventional genome editing tools such as Transcription Activator-Like Effector Nucleases (TALENs) or Zinc Finger Nucleases (ZFNs), CRISPR is a more versatile platform that can be easily adjusted to target new genes of interest.The mechanism behind genome editing by the CRISPR/Cas9 system has been recently thoroughly reviewed elsewhere [1]. Briefly, CRISPR-mediated genome editing is dependent on at least two components: (1) a Cas protein that possesses endonuclease activity and(2) a variable 20 base pair nucleic-acid based targeting crisprRNA (crRNA) that defines the target of interest. Depending on the type of Cas protein employed, also a trans-activating RNA (tracrRNA) is required in order to activate nuclease activity. Together, the gRNA and tracrRNA are often referred to as the single guide RNA, or sgRNA.

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Section: Authors Used a Pooled Crispr Interference Approach To Identifymentioning

confidence: 99%

Combining CRISPR with Flow‐FISH to study CRISPR‐mediated genome perturbation

Freen‐van Heeren

2023

Cytometry Pt A

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“…As discussed, several groups have shown that dysfunctional T cells from cancer patients or murine models express IFNG mRNA, but fail to translate it into IFN‐γ protein [51‐53]. These reports show that post‐transcriptional regulatory mechanisms silence TIL effector function and highlight the importance of assessing mRNA and protein simultaneously in TILs [168]. One of the cis ‐elements that are involved in blocking cytokine production in TILs is AREs.…”

Section: Fine‐tuning Tils: Post‐transcriptional Control Of Anti‐tumour Responsesmentioning

confidence: 99%

Post‐transcriptional control of T‐cell cytokine production: Implications for cancer therapy

Heeren¹

2021

Immunology

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As part of the adaptive immune system, T cells are vital for the eradication of infected and malignantly transformed cells. To perform their protective function, T cells produce effector molecules that are either directly cytotoxic, such as granzymes, perforin, interferon‐γ and tumour necrosis factor α, or attract and stimulate (immune) cells, such as interleukin‐2. As these molecules can also induce immunopathology, tight control of their production is required. Indeed, inflammatory cytokine production is regulated on multiple levels. Firstly, locus accessibility and transcription factor availability and activity determine the amount of mRNA produced. Secondly, post‐transcriptional mechanisms, influencing mRNA splicing/codon usage, stability, decay, localization and translation rate subsequently determine the amount of protein that is produced. In the immune suppressive environments of tumours, T cells gradually lose the capacity to produce effector molecules, resulting in tumour immune escape. Recently, the role of post‐transcriptional regulation in fine‐tuning T‐cell effector function has become more appreciated. Furthermore, several groups have shown that exhausted or dysfunctional T cells from cancer patients or murine models possess mRNA for inflammatory mediators, but fail to produce effector molecules, hinting that post‐transcriptional events also play a role in hampering tumour‐infiltrating lymphocyte effector function. Here, the post‐transcriptional regulatory events governing T‐cell cytokine production are reviewed, with a specific focus on the importance of post‐transcriptional regulation in anti‐tumour responses. Furthermore, potential approaches to circumvent tumour‐mediated dampening of T‐cell effector function through the (dis)engagement of post‐transcriptional events are explored, such as CRISPR/Cas9‐mediated genome editing or chimeric antigen receptors.

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Harnessing high‐throughput flow cytometric protein and RNA detection to enhance TIL therapy

Cited by 2 publications

References 15 publications

Combining CRISPR with Flow‐FISH to study CRISPR‐mediated genome perturbation

Combining CRISPR with Flow‐FISH to study CRISPR‐mediated genome perturbation

Post‐transcriptional control of T‐cell cytokine production: Implications for cancer therapy

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