Product Overview
Parental Cell Line
HCT116
Mutation Description
Knockout achieved by using CRISPR/Cas9, homozygous: 1 bp insertion in exon 2.
Knockout Validation
Sanger Sequencing, Western Blot (WB)
Cell Line Properties
Target Information
Gene Function
May be the important intermediate by which p53/TP53 mediates its role as an inhibitor of cellular proliferation in response to DNA damage. Binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. Functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, inhibits the kinase activity of the cyclin D-CDK4 complex.
Sequence Similarities
Belongs to the CDI family.
Post-translational Modifications
Phosphorylation of Thr-145 by Akt or of Ser-146 by PKC impairs binding to PCNA. Phosphorylation at Ser-114 by GSK3-beta enhances ubiquitination by the DCX(DTL) complex.Ubiquitinated by MKRN1; leading to polyubiquitination and 26S proteasome-dependent degradation. Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, leading to its degradation during S phase or following UV irradiation. Ubiquitination by the DCX(DTL) complex is essential to control replication licensing and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation.
Storage & Handling
Cryopreservation Cell Medium
Cell freezing medium - DMSO serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture Medium
McCoY5a + 10% FBS
Initial Handling Guidelines
Lyophilizate may be stored at 4 °C. After reconstitution, store at -20 °C for short-term storage or -80 °C for long-term storage.
Storage Instructions
Store at -80 °C. Avoid freeze / thaw cycles.
