The AMPD2 knockout HAP1 polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human near-haploid cell line. These cells carry a targeted disruption of the AMPD2 gene, which encodes AMP deaminase 2, and are designed for functional investigation of purine nucleotide metabolism and energy homeostasis. The polyclonal nature of the knockout provides a heterogeneous population suitable for pooled functional screens and robust loss-of-function studies without clone-specific artifacts.
HAP1 cells originate from the chronic myeloid leukemia-derived KBM-7 cell line and maintain a near-haploid karyotype, which facilitates straightforward genetic manipulation and functional annotation. The near-haploid genome ensures that a single CRISPR-mediated disruption effectively silences gene function in the majority of cells, providing an efficient platform for loss-of-function experiments and comparative studies in a diploid-like background during subsequent cell division.
AMPD2 catalyzes the irreversible hydrolytic deamination of AMP to IMP, a committed step in the purine nucleotide cycle that controls the adenylate energy charge and purine biosynthesis. The enzyme is allosterically activated by ATP and inhibited by GTP, while inorganic phosphate stabilizes the enzymatic complex. AMPD2 activity is dynamically regulated by the cellular AMP/ATP ratio, the energy-sensing kinase AMPK, and the hypoxia-inducible factor HIF1A, linking nucleotide metabolism to cellular energy status and oxygen availability. Downstream, IMP generated by AMPD2 serves as a precursor for adenylosuccinate synthetase (ADSS) and adenylosuccinate lyase (ADSL), which collectively regenerate AMP. AMPD2-mediated IMP production also modulates mTORC1 activity, integrating purine metabolism with growth signaling. The enzyme shares structural and functional homology with AMPD1, with which it may form heteromeric complexes in certain tissues.
Disruption of AMPD2 in HAP1 cells impairs the purine nucleotide cycle, leading to altered adenylate energy charge and dysregulation of AMPK and mTORC1 pathways. This knockout model recapitulates key metabolic disturbances observed in diseases such as pontocerebellar hypoplasia type 9 (PCH9), spastic paraplegia, and muscle metabolic disorders. The HAP1 background, with its well-defined genetic context, allows systematic dissection of AMPD2-dependent metabolic and signaling networks, including the interplay between nucleotide pools and energy homeostasis.
Applications include functional genomics screens to identify genetic interactors of the purine nucleotide cycle, profiling of AMPK signaling under metabolic stress, and drug screening for compounds that modulate AMP deaminase activity or mitochondrial metabolism. The polyclonal population enables robust detection of AMPD2 loss through RT-qPCR or Western blotting, while functional analysis can be performed via AMP deaminase activity assays, ATP/AMP ratio measurements, and phospho-AMPK immunoblotting. Cell viability assays under conditions of energy deprivation or hypoxia further validate metabolic vulnerabilities. These cells also serve as valuable controls in CRISPR screens targeting metabolic pathways. For additional technical specifications, validation data, and ordering details, please contact Ascent Research.
