The AK1 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited cell population in which the AK1 gene has been targeted to create a loss-of-function model. This polyclonal pool, generated by transient Cas9/gRNA delivery and selection, contains a mixture of edited alleles and circumvents the need for single-cell cloning. The heterogeneous nature of the population reduces clonal bias and is well suited for experiments where average gene-disruption effects are of primary interest.
HCT 116 is a human colorectal carcinoma cell line with adherent epithelial morphology, bearing an oncogenic KRAS G13D mutation and microsatellite instability (MSI). These cells are extensively used to investigate oncogenic signaling, drug responses, and metabolic adaptations characteristic of MSI colorectal cancers. Their robust proliferation and genetic tractability make them a reliable platform for studying the consequences of gene knockouts in pathways that intersect with KRAS-driven metabolic reprogramming.
AK1 encodes adenylate kinase 1, which catalyzes the equilibrium reaction 2 ADP ? ATP + AMP, maintaining adenine nucleotide balance and energy charge. Upstream factors including cellular AMP/ATP ratio, HIF1A, and oxidative stress regulate AK1 activity. Downstream, AK1 modulates AMPK signaling, nucleotide biosynthesis, and cell cycle progression. It interacts with adenine nucleotides and cooperates with mitochondrial adenylate kinase AK2. As part of the adenylate kinase family, AK1 connects AMP kinase and nucleotide salvage pathways, underscoring its role in energy and nucleotide homeostasis.
In HCT 116 cells, AK1 disruption impairs adenine nucleotide equilibrium, potentially compromising energy-dependent processes and sensitizing cells to metabolic stress. The KRAS G13D and MSI background of these cells drives altered glycolysis and metabolic flexibility, which may heighten dependence on efficient nucleotide turnover. AK1 knockout may uncover synthetic lethal interactions or therapeutic susceptibilities in colorectal cancer, offering a platform to dissect the interplay between nucleotide metabolism and oncogenic signaling under energetic duress.
AK1 Knockout HCT 116 Polyclonal Cells are suitable for metabolic phenotyping via Seahorse flux analysis, ATP bioluminescence assays, and AMP/ATP ratio measurements. Western blotting for phosphorylated AMPK and ACC reports on AMPK pathway activation, while LC-MS quantification of nucleotide pools reveals metabolic shifts. Cell proliferation assays under nutrient stress or drug treatment facilitate synthetic lethal screening. This model supports research into tumor energy metabolism, nucleotide regulation, and colorectal cancer vulnerabilities. For additional details, contact Ascent Research.
