KRAS-G12D Human ORF cDNA Lentivirus
Human KRAS G12D ORF cDNA lentivirus. Lentiviral particles, packaged with expression vector for G12D mutant of human KRAS proto-oncogene, GTPase (KRAS), transcript variant a. An EF1α promoter drives expression of KRAS G12D fused to a C-terminal V5 tag, and a GFP reporter which is separated from the ORF cDNA by a self-cleaving peptide.
These lentiviral particles allow for easy transduction and high stable expression of the indicated ORF cDNA in mammalian cells. The lentiviral vectors have been optimized to provide high gene expression and reliable genome integration, allowing for easy establishment of long-term, stable cell lines with consistent read out of fluorescent (GFP, RFP) or luminescent (firefly luc) reporters. Transcription of the ORF cDNA fused with C-terminal epitope tag is controlled by either EF1a or CMV promoter, while expression of the reporter or selection marker (puromycin or blasticidin) is separated by a self-cleaving peptide to allow separate translation via the same mRNA transcript. In some constructs, the CMV or EF1a promoter drives expression of cDNA-Tag, while the GFP/RFP reporter and drug selection marker are driven by PGK promoter and separated by a self-cleaving peptide. The particles are capable of transducing a variety of difficult-to-transfect cells, including primary and/or thawed cells, making them an extremely versatile product for studying genes in mammalian systems. The accuracy of ORF cDNA cloning has been confirmed by sequencing and the successful protein expression has been validated by transient transfection.
- Optimized vectors – All LipExoGen ORF cDNA lentiviral particles are produced by the third generation system. They feature novel vectors that have been extensively optimized for high, stable gene expression in mammalian cells, with minimal toxicity, and are ideal for use in sensitive cells such as primary and/or thawed cells. CMV or EF1α drives expression of ORF cDNA fused to C-terminal epitope tag, followed by one or two reporter/drug selection markers (GFP, RFP, firefly Luc, puromycin, blasticidin). Each component is separated by self-cleaving peptides to allow independent protein expression from a single mRNA transcript. Alternatively, CMV or EF1α drives expression of C-terminal tagged ORF cDNA, and human or mouse PGK separately drives expression of reporter (GFP, RFP) and selection marker (puromycin, blasticidin), separated by P2A.
- Versatile readout of gene expression – ORF cDNA contains an epitope tag (e.g. V5, Myc, HA, etc.) fused to the C-terminus, which allows for tag-specific antibodies to be used for purification, or for staining in fixed cells or cell lysates. In live cells, puromycin or blasticidin allows stable cell line generation; transduced cells may also be isolated by FACS sorting based on GFP or RFP. GFP or RFP allows for convenient analysis of ORF cDNA expression by simple fluorescence microscopy, flow cytometry, or other common techniques, without unwanted effects of large fusion protein constructs. For in vivo studies, constructs with luciferase can be used to monitor cell growth by in vivo bioluminescence imaging techniques (e.g. IVIS).
- Easily establish stable cell lines expressing gene of interest – LipExoGen ORF cDNA lentiviral particles are ultra-purified and concentrated to high-titer by PEG precipitation and sucrose gradient centrifugation and are suitable for transduction of primary cells, stem cells, and difficult-to-transfect cells including primary or thawed cells. Transduced cells can be FACS sorted by GFP or RFP, or selected by puromycin or blasticidin, to yield stable cells in about 10 days.
- Multiple applications – LipExoGen ORF cDNA lentiviral particles are supplied in serum-free cell culture medium allowing them to be directly applied in vivo. For optimal stability of the lentivirus in frozen storage, the particles are provided at an ideal concentration (about 200-500 µl per aliquot), but they can be easily concentrated with 10 kDa cutoff columns (Millipore-Sigma Cat# UFC510096) for in vivo use where higher concentrations are desirable, such as for stereotactic brain injections.
- Best value & easy customization – All ORF cDNA lentiviral vectors are expression validated and sequence verified. Together with the previously mentioned advantages and a price that is comparable or less vs the competition, LipExoGen ORF cDNA lentiviral particles are the best way to express and study your gene of interest in human or mouse cells. And, if you don’t find what you’re looking for in our catalog, you can easily customize the promoter, tag, reporter, or selection marker of one of our listed ORF cDNA lentivirus and receive the same quality lentivirus without additional cost in about 10 working days. To modify components of a listed ORF cDNA lentivirus, or to request a custom ORF cDNA lentivirus or plasmid based on your specifications, simply contact us.
|KRAS proto-oncogene, GTPase|
|ORF cDNA Lentiviral Particles|
If you require a modification to one of our products, such as a change in reporter or other vector component, please request a custom order. We provide a variety of fast and efficient services for custom cloning or the production of high-quality custom lentiviral particles on demand, usually for the same or comparable price as the listed item. If you’re unable to find your ORF cDNA of interest in our catalog, or you require a different transcript variant than the one listed, please send us a request using the link below.
|KRAS proto-oncogene, GTPase|
|Alias||KRAS Proto-Oncogene, GTPase; V-Ki-Ras2 Kirsten Rat Sarcoma 2 Viral Oncogene Homolog; Kirsten Rat Sarcoma Viral Oncogene Homolog; GTPase KRas; C-Ki-Ras; K-Ras 2; KRAS2; RASK2; V-Ki-Ras2 Kirsten Rat Sarcoma Viral Oncogene Homolog; Kirsten Rat Sarcoma Viral Proto-Oncogene; Cellular Transforming Proto-Oncogene; Cellular C-Ki-Ras2 Proto-Oncogene; Transforming Protein P21; PR310 C-K-Ras Oncogene; C-Kirsten-Ras Protein; K-Ras P21 Protein; Oncogene KRAS2; C-Ki-Ras2; C-K-RAS; K-RAS2A; K-RAS2B; K-RAS4A; K-RAS4B; C-K-Ras; KI-RAS; Ki-Ras; K-Ras; KRAS1; CFC2; RALD; NS3; OES; NS|
|Gene IDs||HGNC:HGNC:6407 Ensembl:ENSG00000133703 MIM:190070|
|Entrez Gene Summary||“This gene, a Kirsten ras oncogene homolog from the mammalian ras gene family, encodes a protein that is a member of the small GTPase superfamily. A single amino acid substitution is responsible for an activating mutation. The transforming protein that results is implicated in various malignancies, including lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas and colorectal carcinoma. Alternative splicing leads to variants encoding two isoforms that differ in the C-terminal region. [provided by RefSeq, Jul 2008]”|