Comprehensive review of CRISPR-based gene editing: mechanisms, challenges, and applications in cancer therapy

Molecular Cancer, Journal Year: 2024, Volume and Issue: 23(1)

Published: Jan. 9, 2024

Abstract The CRISPR system is a revolutionary genome editing tool that has the potential to revolutionize field of cancer research and therapy. ability precisely target edit specific genetic mutations drive growth spread tumors opened up new possibilities for development more effective personalized treatments. In this review, we will discuss different CRISPR-based strategies have been proposed therapy, including inactivating genes tumor growth, enhancing immune response cells, repairing cause cancer, delivering cancer-killing molecules directly cells. We also summarize current state preclinical studies clinical trials highlighting most promising results challenges still need be overcome. Safety delivery are important therapy become viable option. limitations overcome, such as off-target effects, safety, site. Finally, provide an overview opportunities in future directions development. change landscape research, review aims overcome realize potential.

Language: Английский

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SAMHD1 Functions and Human Diseases

Viruses, Journal Year: 2020, Volume and Issue: 12(4), P. 382 - 382

Published: March 31, 2020

Deoxynucleoside triphosphate (dNTP) molecules are essential for the replication and maintenance of genomic information in both cells a variety viral pathogens. While process dNTP biosynthesis by cellular enzymes, such as ribonucleotide reductase (RNR) thymidine kinase (TK), has been extensively investigated, negative regulatory mechanism pools was recently found to involve sterile alpha motif (SAM) domain histidine-aspartate (HD) domain-containing protein 1, SAMHD1. When active, triphosphohydrolase activity SAMHD1 degrades dNTPs into their 2'-deoxynucleoside (dN) subparts, steadily depleting intercellular pools. The differential expression levels activation states various cell types contributes unique that either aid (i.e., dividing T cells) or restrict nondividing macrophages) consumes dNTPs. Genetic mutations induce rare inflammatory encephalopathy called Aicardi-Goutières syndrome (AGS), which phenotypically resembles infection. Recent publications have identified diverse roles double-stranded break repair, genome stability, stress response through interferon signaling. Finally, series were also reported cancer while why is mutated these remains investigated. Here, we reviewed studies begun illuminating highly virology, immunology, biology.

Language: Английский

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HPV upregulates MARCHF8 ubiquitin ligase and inhibits apoptosis by degrading the death receptors in head and neck cancer

PLoS Pathogens, Journal Year: 2023, Volume and Issue: 19(3), P. e1011171 - e1011171

Published: March 3, 2023

The membrane-associated RING-CH-type finger ubiquitin ligase MARCHF8 is a human homolog of the viral ligases Kaposi’s sarcoma herpesvirus K3 and K5 that promote host immune evasion. Previous studies have shown ubiquitinates several receptors, such as major histocompatibility complex II CD86. While papillomavirus (HPV) does not encode any ligase, oncoproteins E6 E7 are known to regulate ligases. Here, we report expression upregulated in HPV-positive head neck cancer (HNC) patients but HPV-negative HNC compared normal individuals. promoter highly activated by HPV oncoprotein E6-induced MYC/MAX transcriptional activation. knockdown cells restores cell surface tumor necrosis factor receptor superfamily (TNFRSF) death FAS, TRAIL-R1, TRAIL-R2, enhances apoptosis. protein directly interacts with TNFRSF receptors. Further, knockout mouse oral expressing HPV16 augments apoptosis suppresses growth vivo. Our findings suggest inhibits upregulating degrading receptors cells.

Language: Английский

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The membrane-associated ubiquitin ligase MARCHF8 stabilizes the human papillomavirus oncoprotein E7 by degrading CUL1 and UBE2L3 in head and neck cancer

Journal of Virology, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 16, 2024

ABSTRACT The human papillomavirus (HPV) oncoprotein E7 is a relatively short-lived protein required for HPV-driven cancer development and maintenance. degraded through ubiquitination mediated by cullin 1 (CUL1) the ubiquitin-conjugating enzyme E2 L3 (UBE2L3). However, proteins are maintained at high levels in most HPV-positive cells. A previous proteomics study has shown that UBE2L3 CUL1 increased knockdown of E3 ubiquitin ligase membrane-associated ring-CH-type finger 8 (MARCHF8). We have recently demonstrated HPV16 upregulates MARCHF8 expression keratinocytes head neck (HPV+ HNC) Here, we report stabilizes degrading components S-phase kinase-associated 1-CUL1-F-box complex HPV+ HNC found cells drastically decreases level while increasing levels. further revealed binds to ubiquitinates enhances protein. Conversely, overexpression suppresses tumor growth vivo . Our findings suggest HPV-induced prevents degradation ubiquitinating proteins. IMPORTANCE Since essential virus replication; HPV maintain HPV-infected can be efficiently ubiquitinated proteasomes host cell. Mechanistically, (UBE2L3) play an role degradation. show membrane (MARCHF8) induced E6 blocking proteasomes. knockout restores expression, decreasing inhibiting proliferation Additionally, or growth. results maintains cell inducing MARCHF8, which may critical tumorigenesis.

Language: Английский

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SAMHD1 deacetylation by SIRT1 promotes DNA end resection by facilitating DNA binding at double-strand breaks

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: Nov. 7, 2022

Abstract Sterile alpha motif and HD domain-containing protein 1 (SAMHD1) has a dNTPase-independent function in promoting DNA end resection to facilitate double-strand break (DSB) repair by homologous recombination (HR); however, it is not known if upstream signaling events govern this activity. Here, we show that SAMHD1 deacetylated the SIRT1 sirtuin deacetylase, facilitating its binding with ssDNA at DSBs, promote HR. complexes deacetylates conserved lysine 354 (K354) specifically response DSBs. K354 deacetylation promotes HR but tetramerization or dNTPase Mechanistically, recruitment DSBs which turn facilitates CtIP binding, leading promotion of genome integrity. These findings define mechanism governing stability.

Language: Английский

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A Critical Role for p53 during the HPV16 Life Cycle

Microbiology Spectrum, Journal Year: 2022, Volume and Issue: 10(3)

Published: May 24, 2022

Human papillomaviruses (HPV) are causative agents in ano-genital and oral cancers; HPV16 is the most prevalent type detected human cancers. The E6 protein targets p53 for proteasomal degradation to facilitate proliferation of infected cell. However, immortalized cells predominantly spliced (E6*) unable degrade p53. Here, we demonstrate that foreskin keratinocytes by (HFK+HPV16), positive oropharyngeal cancers, retain significant expression In addition, levels increase HPV16+ head neck cancer cell lines following treatment with cisplatin. Introduction full-length into HFK+HPV16 resulted attenuation cellular growth (in hTERT HFK, promoted enhanced proliferation). An understudied interaction between E2 investigated whether this was important viral life cycle. We generated mutant genomes interact resulting profound phenotypes primary HFK. induced hyper-proliferation, but an ultimate arrest growth; β-galactosidase staining demonstrated increased senescence, COMET assays showed DNA damage compared wild-type cells. There failure cycle organotypic rafts HFK premature differentiation reduced proliferation. results critical during cycle, may be due a functional Disruption has antiviral potential.

Language: Английский

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SAMHD1 … and Viral Ways around It

Viruses, Journal Year: 2021, Volume and Issue: 13(3), P. 395 - 395

Published: March 2, 2021

The SAM and HD domain-containing protein 1 (SAMHD1) is a dNTP triphosphohydrolase that plays crucial role for variety of different cellular functions. Besides balancing intracellular concentrations, facilitating DNA damage repair, dampening excessive immune responses, SAMHD1 has been shown to act as major restriction factor against various virus species. In addition its well-described activity retroviruses such HIV-1, identified reduce the infectivity viruses herpesviruses CMV EBV, poxvirus VACV, or hepadnavirus HBV. While some are efficiently restricted by SAMHD1, others have developed evasion mechanisms antagonize antiviral SAMHD1. Within this review, we summarize functions highlight countermeasures evolved neutralize

Language: Английский

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SAMHD1 enhances HIV-1-induced apoptosis in monocytic cells via the mitochondrial pathway

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 10, 2025

SUMMARY Sterile alpha motif (SAM) and histidine-aspartate (HD) domain-containing protein 1 (SAMHD1) inhibits HIV-1 replication in non-dividing cells by reducing the intracellular dNTP pool. SAMHD1 enhances spontaneous apoptosis cells, but its effects on HIV-1-induced underlying mechanisms remain unknown. Here we uncover a new mechanism which monocytic through mitochondrial pathway. We found that endogenous levels induced infection dividing THP-1 cells. Mechanistically, expression decreases membrane potential promotes cytochrome c release thereby enhancing apoptotic SAMHD1-enhanced is associated with increased of pro-apoptotic BCL-2-interacting killer (BIK) further demonstrated BIK contributes to during infection. Overall, our results reveal an unappreciated regulatory via pathway

Language: Английский

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dNTP depletion and beyond: the multifaceted nature of SAMHD1-mediated viral restriction

Journal of Virology, Journal Year: 2025, Volume and Issue: unknown

Published: April 25, 2025

ABSTRACT SAMHD1 is a dNTPase of mammalian cells. In 2011, was found to be host restriction factor against retroviruses through dNTP reduction. Recent research provides evidence that the antiviral mechanisms cannot explained solely by its activity. Instead, versatility SAMHD1-mediated various viruses suggests extend beyond depletion. This explains multifaceted and broad functions play significant role in innate immunity.

Language: Английский

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Werner Syndrome Protein (WRN) Regulates Cell Proliferation and the Human Papillomavirus 16 Life Cycle during Epithelial Differentiation

mSphere, Journal Year: 2020, Volume and Issue: 5(5)

Published: Sept. 15, 2020

Human papillomaviruses recruit a host of DNA damage response factors to their viral genome facilitate homologous recombination replication in association with the E1 and E2. We previously demonstrated that SIRT1 deacetylation WRN promotes recruitment E1-E2 replicating regulates both levels fidelity replication. The by results an active protein able complex DNA, but is less stable. Here, we demonstrate inverse correlation between CIN cervical lesions compared normal control tissue, supporting our model destabilizing protein. CRISPR/Cas9 edited N/Tert-1 N/Tert-1+HPV16 cells knock out expression subjected organotypic raft cultures. In without expression, there was enhanced basal cell proliferation, damage, thickening differentiated epithelium. cells, increased throughout epithelium, Overall, required proliferation controls HPV16 life cycle these cells. This complements previous data demonstrating describe new role for WRN, tumor suppressor, controlling keratinocyte differentiation cycle.

Language: Английский

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