The JMY Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the JMY gene in the NCI-H1975 human lung adenocarcinoma cell line. This loss-of-function model is generated through CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous pool of edited cells suitable for studying JMY-dependent cellular processes without clonal selection. The polyclonal format preserves the biological variability of the knockout, enabling robust functional analyses in a genotypically diverse population. Researchers can utilize these cells to dissect JMY??s dual roles in transcriptional regulation and cytoskeletal dynamics.
The NCI-H1975 host cell line is a well-characterized epithelial cell model derived from the pleural effusion of a female patient with non-small cell lung adenocarcinoma. These cells harbor endogenous EGFR L858R and T790M mutations, conferring sensitivity and acquired resistance to first-generation EGFR tyrosine kinase inhibitors, and are widely employed in investigations of EGFR-targeted therapeutic resistance. The genetic background of NCI-H1975 also features alterations in p53 pathway components, making it a valuable system for examining p53-dependent and -independent cell fate decisions in a clinically relevant context.
JMY functions as a transcriptional cofactor for p53 (TP53), augmenting the expression of p53 target genes such as CDKN1A (p21), BAX, and PUMA, thereby facilitating cell cycle arrest and DNA damage-induced apoptosis. Upstream, JMY is regulated by DNA damage sensors including ATM and ATR in response to genotoxic stress. Beyond its nuclear role, JMY operates in the cytoplasm as a regulator of actin nucleation, interacting with WASF3 and the Arp2/3 complex (including ARPC1A) to promote F-actin polymerization and influence cell migration. Through its scaffold protein STRAP, JMY integrates signals from the DNA damage response and cytoskeletal reorganization. Representative pathway components linking JMY to these processes include TP53, JMY, CDKN1A, BAX, WASF3, ARPC2, ACTB, and CFL1.
In the NCI-H1975 background, knockout of JMY disrupts a critical node connecting p53-mediated transcription and actin cytoskeleton regulation, offering a tool to interrogate how loss of p53 cofactor function impacts non-small cell lung cancer cell behavior. Given the prevalence of p53 pathway dysregulation and EGFR TKI resistance in NSCLC, this model is particularly suited for examining the interplay between DNA damage repair mechanisms, apoptotic threshold, and metastatic potential. The polyclonal knockout population permits the study of heterogeneous responses to therapeutic agents, reflecting the intratumoral diversity encountered in clinical settings.
This knockout model supports Western blotting for p53 targets, RT-qPCR for JMY-regulated genes, and immunofluorescence for F-actin. Functional assays include wound healing, Transwell migration, and flow cytometry for apoptosis and cell cycle. Co-immunoprecipitation of p53-JMY and ChIP-qPCR for p53 binding sites are feasible. Drug sensitivity testing with cisplatin or etoposide further extends applications. For specifications, contact Ascent Research.
