The AKR1C3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 cell line, generated through targeted disruption of the AKR1C3 gene. This loss-of-function model abolishes the enzymatic activity of AKR1C3, a critical hydroxysteroid dehydrogenase responsible for the final step in androgen and estrogen biosynthesis, as well as prostaglandin F2?? production. The polyclonal nature of the product captures a broad spectrum of gene-editing outcomes, providing a robust population-level tool for functional genomics without the biases of clonal isolation.
HAP1 is a chronic myeloid leukemia (CML)-derived cell line with a stable near-haploid karyotype, originally isolated from the KBM-7 lineage of a 39-year-old male in blast crisis. Its haploid genetic constitution makes it particularly advantageous for CRISPR/Cas9 experiments, as disruption of a single allele tends to yield a complete loss-of-function phenotype. This feature, combined with reliable growth kinetics and extensive characterization, has established HAP1 as a workhorse for genetic screens, drug sensitivity profiling, and mechanistic studies in cancer biology.
AKR1C3 functions as an aldoketo reductase that catalyzes the NADPH-dependent conversion of androstenedione to testosterone and estrone to estradiol, directly fueling androgen receptor (AR) and estrogen receptor (ER) signaling pathways. In parallel, it reduces prostaglandin H2 to prostaglandin F2?? (PGF2??), which activates the PTGFR receptor to modulate inflammatory and proliferative responses. Transcriptionally, AKR1C3 is regulated by AR, ER, Nrf2, NF-??B, and IL-6/STAT3, often in response to oxidative stress. Its activity promotes the expression of AR target genes (e.g., PSA, TMPRSS2) and ER targets (e.g., PGR, GREB1), while also engaging PI3K/AKT and MAPK/ERK downstream cascades, thereby integrating hormonal signaling with growth and survival pathways.
In the HAP1 context, AKR1C3 knockout is predicted to diminish intracellular testosterone and estradiol pools, reduce PGF2?? synthesis, and attenuate downstream AR/ER and PTGFR-dependent transcriptional programs. This can recalibrate cellular proliferation, differentiation, and apoptotic thresholds, and may alter sensitivity to hormone-directed therapies such as enzalutamide or abiraterone. The haploid background ensures high penetrance of the gene disruption, making this polyclonal pool a reliable and scalable model for interrogating AKR1C3 biology in a leukemia-derived, hormone-sensitive framework.
This polyclonal knockout pool is ideally suited for validating AKR1C3 inhibitors, profiling steroid hormone levels via LC-MS/MS, conducting AR/ER reporter assays, and assessing drug sensitivity in proliferation or apoptosis assays. It also provides a robust negative control for kinome- or metabolome-wide CRISPR screens targeting steroidogenic networks, and is compatible with transcriptomic analysis by RNA-seq. For additional information on product validation, culture conditions, and ordering, please contact Ascent Research.
