The KLHL26 Knockout HCT 116 Polyclonal Cells provide a CRISPR/Cas9-mediated gene disruption model in the KLHL26 gene, delivered as a polyclonal knockout cell population. Unlike monoclonal derivatives, this format encompasses a spectrum of edit types, offering built-in biological replication and reducing the risk of clonal artifacts. It is particularly well-suited for studies requiring bulk cellular behavior, pooled phenotypic screens, or as a source for further clonal isolation if needed.
The host HCT 116 cell line is a human colorectal carcinoma model originating from a male patient. Its genomic profile includes an MLH1 mutation conferring microsatellite instability-high (MSI-H), an activating KRAS G13D mutation, and wild-type TP53. These characteristics render HCT 116 cells a standard system for investigating mismatch repair-deficient colorectal cancer, oncogenic signaling, and inherent resistance or sensitivity to chemotherapeutics.
KLHL26 functions as a substrate recognition component of the CUL3-RBX1 E3 ubiquitin ligase complex, which catalyzes ubiquitination and proteasomal degradation of YAP1, a key transcriptional coactivator of the Hippo pathway. Through this mechanism, KLHL26 serves as a negative regulator of Hippo signaling, counteracting the pro-proliferative and pro-migratory activities of YAP1 and its paralog TAZ. Upstream regulators include the Hippo pathway kinases MST1/2 and LATS1/2, which modulate KLHL26 function, as well as oxidative stress. Upon KLHL26-mediated YAP1 destruction, TEAD-dependent transcription is suppressed, decreasing expression of target genes such as CTGF and CYR61. Thus, KLHL26 links the ubiquitin-proteasome system to Hippo pathway control, impacting cell cycle progression and cytoskeletal dynamics.
In the HCT 116 background, loss of KLHL26 is anticipated to elevate YAP1/TAZ activity, potentially driving enhanced TEAD-mediated transcription and synergistic effects with the existing KRAS G13D oncogenic mutation. This knockout model enables dissection of how Hippo pathway components interact with MAPK signaling and genomic instability. It is an invaluable tool for studying the role of YAP1/TAZ in colorectal cancer initiation, progression, and metastasis, particularly in the context of defective DNA mismatch repair and mutant RAS.
Researchers can apply these cells in numerous assays: western blotting to monitor total and phosphorylated YAP1; RT-qPCR for downstream targets (CTGF, CYR61); co-immunoprecipitation to analyze KLHL26-CUL3 binding; and in vitro ubiquitination assays to assess YAP1 modification. Functional readouts include cell proliferation, migration, invasion, and colony formation assays. Immunofluorescence can visualize YAP1 subcellular translocation, and the cells are suitable for xenograft tumor models. Applications span Hippo pathway investigation, ubiquitin-proteasome biology, and colorectal cancer research, including drug target identification and validation. For technical inquiries, please contact Ascent Research.
