Product Overview
Parental Cell Line
HEK293T
Mutation Description
Knockout achieved by using CRISPR/Cas9, homozygous: 180 bp insertion in exon 2.
Knockout Validation
Sanger Sequencing, Western Blot (WB)
Cell Line Properties
Target Information
Gene Function
Kinase subunit of both mTORC1 and mTORC2, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-421', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Phosphorylates MAF1 leading to attenuation of its RNA polymerase III-repressive function. mTORC2 is also activated by growth. factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. mTORC2 plays a critical role in AKT1 'Ser-473' phosphorylation, which may facilitate the phosphorylation of the activation loop of AKT1 on 'Thr-308' by PDK1 which is a prerequisite for full activation. mTORC2 regulates the phosphorylation of SGK1 at 'Ser-422'. mTORC2 also modulates the phosphorylation of PRKCA on 'Ser-657'.
Sequence Similarities
Belongs to the PI3/PI4-kinase family. Contains 1 FAT domain. Contains 1 FATC domain. Contains 7 HEAT repeats. Contains 1 PI3K/PI4K domain.
Post-translational Modifications
Autophosphorylated; when part of mTORC1 or mTORC2.
Storage & Handling
Cryopreservation Cell Medium
Cell freezing medium - DMSO serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture Medium
DMEM (High Glucose) + 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.
