Table 3 Bioengineered cancer cell lines using CRISPR technology

Murine melanoma B16F10 cell line

• CD47: It acts as a signal on tumor cells to prevent their phagocytosis by macrophages

• IL-12: Promotes immune response against tumors by inhibiting angiogenesis, boosting T-cell growth, and inducing TAM polarization

• Tumor cells were genetically engineered to produce IL-12 while simultaneously knocking out the CD47 gene

• This dual modification enhanced the immune response by promoting increased macrophage activity

[240]

Human melanoma cell lines WM115, CM150-Post, and NZM40

• Early B-cell factor 3 (EBF-3): It acts as a putative epigenetic driver of melanoma metastasis

• DNA methylation editing was conducted, with particular emphasis on the EBF-3 promoter region

• Study highlights the promising capabilities of CRISPR for conducting epigenetic editing in disease-related investigations

[241]

The human NSCLC cell line A549 and the cisplatin-resistant

A549 cell line (A549/DDP)

• NNT: It encodes for an enzyme known as nicotinamide nucleotide transhydrogenase, which plays a crucial role in mitochondrial energy metabolism

• CRISPR-based DNA hypermethylation leads to the silencing of NNT in cisplatin-resistant lung cancer cells

• Restoring NNT expression reduces resistance to cisplatin and suppresses autophagy

[242]

ID8 cells (murine ovarian surface epithelial cells)

• Trp53/p53: A crucial tumor suppressor gene that is vital in controlling the cell cycle

• Breast cancer susceptibility gene 2: Functions as a tumor suppressor gene, aiding in the prevention of cancer cell formation

• Novel ID8 derivatives were generated by introducing single or double deletions of suppressor genes

• This approach enabled the modeling of high-grade serous carcinoma for further study

[243]

NPC (Nasopharyngeal carcinoma) cell line CNE2

• KLHDC4: It interacts with tumor suppressor protein p53, thereby augmenting its capacity to initiate apoptosis in cancer cells

• Gene editing of KLHDC4 led to the inhibition of cell proliferation, reduced colony formation, and suppressed tumor growth in mice

• Findings suggest KLHDC4 holds promise as a potential therapeutic target and a prognostic biomarker

[244]

SH-Y5Y neuroblastoma cell line

• Chromosome 6q region: Loss of 6q genetic material, including tumor suppressor CDKN1A, affects cell growth and proliferation and is implicated in neuroblastoma progression

• Chromosome 11q region: 11q genetic material loss, including tumor suppressors ATM and CBL, is linked to aggressive neuroblastoma

• CRISPR‒Cas9 mediated deletions in chromosome 11q and 6q revealed oncogenic advantages, suggesting therapeutic potential in targeting these deletions

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Human cell lines Panc-1 and SUIT-2 and the murine cell line TB32047

• KrasG12D: It is postulated to be a key driver in tumor initiation and progression

• Three Kras heterozygous cell lines were investigated, and the resulting knockout clones displayed characteristics akin to those of wild-type cells during standard growth conditions

[246]

Adriamycin-resistant (A2780/ADR) ovarian cancer cell line

• ABCB1: It encodes P-glycoprotein (a membrane protein that aids in the transport of molecules across cell membranes). ABCB1 mutations are associated with anticancer drug resistance

• Downregulation of the ABCB1 results in heightened sensitivity of drug-resistant ovarian cancer cells to doxorubicin treatment

[247]

PC-9 (lung adenocarcinoma cell line)

• Epidermal growth factor receptor (EGFR): It encodes a protein that governs cell growth, survival, and migration. Abnormalities in this gene can drive cancer growth

• CRISPR/Cas9 introduced EGFR T790M mutation into PC9 lung cancer cells, creating clones resistant to gefitinib but sensitive to AZD9291

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