pLenti-EEF2K-sgRNA

Cat. No.: P745533

Specifications

Product Name

pLenti-EEF2K-sgRNA

Product Line

Produtos biológicos

Armazenamento e envio

Condições de armazenamento de armazenamento

Store at -80°C

Enviado em

Dry ice packs + Cold packs

Estabilidade e armazenamento

Stored at -20 ℃, valid for 3 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

No tested applications or protocols are provided in the Product Data for this SKU. Recommended use cases (e.g., transfection, transduction, selection parameters, or assay workflows) are not specified for this item; refer to the CoA/Spec Sheet and SDS. Any experimental implementation should follow institutional biosafety policies and standard operating procedures for genetic constructs.

Biological Roles

Target context (general, literature-level):

• EEF2K (eukaryotic elongation factor 2 kinase) is an atypical, Ca2+/calmodulin‑dependent protein kinase that phosphorylates eEF2 on Thr56, reducing its affinity for the ribosome and slowing translation elongation.
• EEF2K integrates nutrient and stress signals, downstream and in crosstalk with pathways such as AMPK and mTORC1. Under energy stress or limited nutrients, EEF2K activation conserves resources by dampening elongation.
• Regulation includes calmodulin binding, phosphorylation by upstream kinases (e.g., S6K, p90RSK in literature), and feedback from cellular energy status.
• Cellular processes influenced: Global protein synthesis rate, autophagy regulation, and adaptation to hypoxia or oxidative stress have been associated with EEF2K activity in multiple model systems.

Relevance to this product:

• An sgRNA directed to EEF2K enables perturbation of this regulatory node, facilitating studies of translational control, stress signaling, and metabolic adaptations. Specific target site(s), spacer sequence, and species for this item are not provided in the Product Data and should be confirmed from the CoA/Spec Sheet before experimental use.

Buffer Applications

Not typically applicable. This product is a genetic construct, not a buffering agent. Any supplied storage or dilution buffer details are not specified in the Product Data; refer to the CoA/Spec Sheet for composition if needed.

Green Alternatives

Environmental considerations for nucleic acid reagents focus on ancillary materials rather than the construct itself.

• DNA/RNA handling typically avoids hazardous organic solvents; aqueous, nuclease‑free buffers predominate.
• For associated workflows, consider:
  • Using room‑temperature stable enzymes/reagents where feasible to reduce cold‑chain energy demands (trade‑off: may not match performance of cold‑stored reagents).
  • Selecting non‑toxic, biodegradable decontamination agents for bench and biosafety cabinet cleaning (trade‑off: efficacy and contact times vary).
  • Minimizing single‑use plastics by adopting validated low‑waste consumables (trade‑off: potential risk of cross‑contamination if re‑use is attempted without validated sterilization).

Comparison (general):

• Conventional approach: Cold‑chain intensive storage/shipping (−20 to −80 °C), single‑use plastics, bleach for decontamination.
• Greener approach: Optimized inventory management to reduce reships; energy‑efficient ultra‑low freezers; validated alternative disinfectants; consolidation of shipments.

Note: No item‑specific green certification or alternative formulation is provided in the Product Data.

Pharmaceutical Uses

Not applicable. This product is designated For research use only and is not intended for diagnostic, therapeutic, or manufacturing use in humans or animals. No pharmacopeial status or excipient role is provided for this item.

Physical Properties

As a nucleic acid construct, conventional small‑molecule physical constants are not applicable.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration/solvent/buffer: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight/formula: Not specified; plasmid/construct dependent.
• MP/BP/density/refractive index/logP/pKa: Not applicable to DNA constructs; not specified for this item.
• Solubility: DNA/RNA constructs are typically supplied in aqueous buffer; the exact solvent system for this item is not specified; refer to CoA/Spec Sheet.

Notes (general, literature-level):

• Nucleic acids are water-soluble under physiological ionic strength and are stabilized by low-salt, nuclease-free buffers. Handling on ice and minimizing freeze–thaw cycles help preserve integrity, but the precise buffer and stabilizers for this product are not provided.

Quality and Grades

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

Context for nucleic acid reagent quality (general):

• Sequence identity and integrity are typically verified by restriction analysis and/or Sanger/NGS confirmation; for functional sgRNA vectors, spacer sequence verification is critical.
• Endotoxin and host‑cell DNA/RNA/protein carryover can affect cell‑based studies; plasmid prep grade (e.g., research grade vs. transfection grade) determines typical endotoxin limits and sterility controls. Exact acceptance criteria for this item are not specified.
• Sterility: For cell culture applications, reagents are commonly filtered and produced under clean conditions; specific sterility testing for this item is not provided.
• Functional validation: sgRNA activity is often validated by indel formation at the intended locus; no functional data are provided in the Product Data.

Please refer to the lot‑specific Certificate of Analysis/Specification Sheet for definitive quality attributes (concentration, sequence, size, endotoxin, sterility, and acceptance criteria) applicable to this SKU.

Reaction and Applications

This product name indicates a pLenti vector encoding an sgRNA targeting EEF2K, suited for CRISPR applications delivered via lentiviral backbones.

Applications (general, literature-level; no protocol specifics):

• Gene function studies: Enable loss‑of‑function interrogation of EEF2K by directing CRISPR nuclease systems to the EEF2K locus for indel generation and frameshift knockouts in dividing and non‑dividing cells following stable delivery.
• Stable cell line generation: Lentiviral delivery supports integration and long‑term sgRNA expression, facilitating durable gene perturbation and downstream phenotypic assays.
• Pathway mapping: EEF2K regulates translation elongation via phosphorylation of eEF2; perturbation informs studies on translational control, stress responses, and nutrient‑sensing pathways.
• Complementary modalities: Can be paired with catalytically inactive CRISPR effectors (e.g., KRAB for CRISPRi or activators for CRISPRa) when the sgRNA is compatible with those systems.

Implementation notes (conceptual only):

• Validate on‑target editing at EEF2K with orthogonal genotyping and functional readouts.
• Assess potential off‑target effects using guide design best practices and appropriate controls.
• Use institutional biosafety and quality frameworks for handling lentiviral constructs. Specific conditions, sequences, or procedures for this item are not provided in the Product Data.

Reaction Conditions

Actionable procedural details are not provided in the Product Data. The following are high‑level, literature‑based considerations without stepwise instructions:

• Delivery modality: Lentiviral backbones are commonly used for stable integration across a broad range of cell types, including those with low division rates.
• System components: The sgRNA requires a compatible CRISPR effector (e.g., Cas9) expressed in the same cells. Expression source (integrated vs. transient) influences kinetics and durability of editing.
• Verification: On‑target activity is typically evaluated via genomic assays (e.g., indel detection) and functional readouts relevant to EEF2K pathway modulation.
• Controls: Include non‑targeting sgRNA, known on‑target positive controls, and rescue constructs as appropriate to disentangle on‑target from off‑target and delivery effects.

No specific temperatures, times, multiplicities, sequences, or reagent ratios are provided for this SKU. Consult your institution’s biosafety guidance and the item’s CoA/Spec Sheet for any lot‑specific recommendations.

Safety and Handling

• GHS classification, signal word, pictograms, and H‑statements: Not specified for this item; refer to the SDS.
• Research use note: For research use only.
• Storage conditions: Store at −80 °C (per Product Data). Avoid repeated freeze–thaw.
• Shipping: Dry ice packs + cold packs (per Product Data) to maintain frozen/cold chain.

General guidance (non‑exhaustive; defer to institutional biosafety and SDS):

• This item is a genetic construct. Follow institutional biosafety policies appropriate for handling recombinant DNA/viral vectors. The specific biosafety level depends on formulation and local risk assessment.
• Use standard laboratory PPE (lab coat, gloves, eye protection). Work in a clean, nuclease‑free environment to prevent degradation and cross‑contamination.
• Incompatibilities: Avoid nucleases and non‑sterile equipment. Keep away from elevated temperatures and prolonged light exposure that may impact nucleic acid integrity (general caution).
• First aid (overview): In case of accidental exposure, follow routine laboratory first‑aid measures and consult the SDS and institutional guidelines. Seek medical attention as required.

Always consult the SDS provided with the shipment for authoritative hazard and handling information.

Solvent Selection

Not typically applicable. This is a nucleic acid construct rather than a small‑molecule solute.

Practical note (general):

• Such constructs are usually provided in nuclease‑free aqueous buffers. If dilution is required, nuclease‑free water or the supplier‑specified buffer is typically used to maintain stability. For this item, the exact buffer system is not specified; consult the CoA/Spec Sheet.

Storage and Reconstitution

• Storage conditions: Store at −80 °C (per Product Data).
• Shipping: Dry ice packs + cold packs to maintain cold chain (per Product Data).
• Thawing/reconstitution buffer, concentration, aliquoting guidance: Not specified for this item; refer to CoA/Spec Sheet.
• Shelf life/expiry: Not specified for this item; refer to CoA/Spec Sheet.

General notes (non‑specific): Maintain nuclease‑free technique and minimize freeze–thaw to preserve nucleic acid integrity. Follow any lot‑specific instructions provided with the shipment.

Structure and Identity

This product is a lentiviral CRISPR sgRNA construct designated to target the EEF2K gene (eukaryotic elongation factor 2 kinase) within a pLenti backbone. It is a biological construct (nucleic acid-based), not a small molecule.

• Product name: pLenti-EEF2K-sgRNA (SKU: P745533)
• Construct type: Lentiviral sgRNA expression vector (general description; specific backbone map and promoter elements are not provided in the Product Data)
• Target: EEF2K (gene symbol EEF2K)
• Molecular formula: Not applicable for a plasmid/construct; not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES/InChI/InChIKey/CID/CAS: Not applicable to nucleic acid constructs; not specified for this item.

Structural features (general, literature-level):

• Typical pLenti sgRNA systems encode an RNA polymerase III promoter (e.g., U6) driving a single-guide RNA scaffold; the exact elements for this item are not specified.
• The 2D “structure” is a plasmid map (circular DNA) rather than a defined molecular structure; sequence-level features (sgRNA spacer sequence, selectable markers, packaging signals) are not provided in the Product Data.

Synthetic Utility

In the context of genome engineering rather than chemical synthesis, the “utility” pertains to genetic perturbation workflows.

• Functional group/element: The sgRNA spacer and scaffold drive sequence‑specific DNA recognition by a CRISPR nuclease supplied by the user’s system. The pLenti backbone supports stable delivery.
• Use in combinatorial studies: Multiple sgRNAs can be multiplexed to interrogate redundancy or generate larger deletions (conceptually). This item’s compatibility with multiplexing is not specified and depends on its backbone and cloning strategy.
• Retrosynthetic value (conceptual): Targeting EEF2K enables upstream or downstream pathway dissection. For example, perturbation of EEF2K can be paired with reporters of translation elongation or phospho‑status of eEF2 to map causal links.
• Complementary tools: Rescue experiments using sgRNA‑resistant EEF2K transgenes or kinase‑dead mutants can clarify on‑target phenotypes. These rescue constructs and exact sgRNA target site(s) are not provided in the Product Data and should be defined by the user based on the supplied sequence.

Target Specificity

• Designation: pLenti-EEF2K-sgRNA (from Product Name)
• Intended target: EEF2K (eukaryotic elongation factor 2 kinase)
• Species/isoform, spacer sequence, and target exon/locus details: Not specified for this item; refer to CoA/Spec Sheet.
• Backbone/context: pLenti sgRNA construct (general description based on name); exact vector map, promoters, selection markers, and any fluorescent reporters are not specified for this item.

Only the information above, derivable from the Product Data, is provided. Sequence‑level specificity and validated off‑target profiles are not included and should be confirmed from the lot‑specific documentation.