pLenti-ABCD3-sgRNA

Cat. No.: P743541

Spécifications

Product Name

pLenti-ABCD3-sgRNA

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

Store at -80°C

Expédié en

Dry ice packs + Cold packs

Stabilité et stockage

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

Application Protocols

Not specified for this item; refer to CoA/Spec Sheet and the manufacturer’s instructions.

Note: Because this is a plasmid DNA reagent, standard CRISPR-Cas9 and lentiviral packaging protocols from the literature or institutional SOPs are typically followed. However, per catalog rules, only product-provided, validated protocols should be cited here.

Biological Roles

General biological context (literature; not item-specific claims)

• Gene target: ABCD3 (ATP-binding cassette sub-family D member 3), also known as PMP70, encodes a peroxisomal transporter implicated in importing very-long-chain fatty acyl-CoAs and related substrates into peroxisomes for β-oxidation.
• Cellular localization: Peroxisomal membrane protein with two transmembrane domains and nucleotide-binding domains characteristic of ABC transporters (literature).
• Functional significance: ABCD3 participates in lipid metabolism and peroxisome biogenesis; perturbation can alter peroxisomal β-oxidation flux and cellular lipid homeostasis. It may function redundantly or complementarily with other ABCD family members (ABCD1/2) depending on substrate specificity (literature).
• Relevance of sgRNA targeting: CRISPR disruption of ABCD3 enables mechanistic studies of peroxisomal transport, metabolic rewiring, and compensatory pathways. Phenotypes are assay- and model-dependent and should be empirically validated.

Notes

• The above describes biological roles of the endogenous gene/protein. It does not specify performance of this particular sgRNA construct, which must be confirmed experimentally (on-target efficacy, off-target profile, and cellular context).

Buffer Applications

Not a classical buffer reagent; however, plasmid DNA handling relies on appropriate aqueous buffers (general guidance; not item-specific specs):

• Storage/short-term handling: Nuclease-free water or TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) supports DNA stability; EDTA chelates divalent cations that catalyze nuclease activity. For transfection, many users dilute in nuclease-free water to avoid EDTA carryover.
• Electrophoresis: Use TAE or TBE for agarose gels when checking plasmid integrity; include safer DNA stains where possible.
• Transfection complexing: Follow reagent manufacturer’s buffer recommendations (often serum-free, low-salt media during complex formation). Excess salt can reduce transfection efficiency.

Item-specific buffer composition, if any, is Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

While “green alternatives” are typically discussed for organic solvents, nucleic-acid workflows also benefit from safer, lower-impact choices. The following are general, literature-based considerations (not item-specific specs):

• Safer nucleic-acid stains: Prefer SYBR Safe or GelGreen over ethidium bromide to reduce mutagenic risk while maintaining sensitivity.
• Buffer choices: Use minimal effective volumes of aqueous, nuclease-free buffers; avoid unnecessary chaotropes and organic extractions when column-based cleanup suffices.
• Packaging/transfection reagents: Consider biodegradable or less cytotoxic transfection reagents to minimize hazardous waste and improve cell health.
• Decontamination: Use UV or hypochlorite judiciously; avoid excessive use of aldehyde-based sterilants.

Illustrative comparison (general)

• Ethidium bromide vs SYBR Safe: Similar sensitivity for gel imaging; SYBR Safe is less hazardous but may have slightly different excitation/emission requirements and cost considerations.
• Phenol–chloroform extraction vs silica spin columns: Spin columns reduce solvent waste and exposure, albeit with consumable plastic use and potential lower recovery for very large plasmids.

Trade-offs

• “Greener” alternatives may involve higher consumable costs or instrumentation needs (e.g., blue-light transilluminators). Balance lab safety, waste reduction, and data quality.

Pharmaceutical Uses

Not applicable as a formulated pharmaceutical or excipient. This product is a research-use-only plasmid DNA reagent for laboratory studies (per Product Data: For research use only). It is not intended for human or veterinary use, clinical diagnostics, or therapeutic applications.

General industry context (literature; not item-specific)

• Lentiviral plasmids and sgRNA cassettes are commonly used in discovery research and preclinical process development to generate edited cell lines or to study gene function. Any use in GMP environments would require purpose-built materials, full documentation, and compliance with regulatory and biosafety requirements beyond the scope of this research reagent.

Physical Properties

Item-specific facts (from Product Data)

• Appearance, concentration, solvent/buffer, plasmid length (bp), antibiotic resistance, copy number: Not specified for this item; refer to CoA/Spec Sheet.

General/literature values (context only; not product specs)

• Nominal mass of dsDNA: ~660 g/mol per base pair (literature approximation for calculating plasmid molecular mass from length, if known).
• Solubility: Plasmid DNA is water-soluble in nuclease-free water or low-salt buffers (e.g., TE); stability enhanced by Tris/EDTA at neutral pH (literature).
• pH stability: DNA is most stable near pH 7–8; hydrolyzes more rapidly under strong acid or base and at elevated temperatures (literature).
• Optical properties: A260/A280 ~1.8 indicates relatively pure DNA; A260 of 1.0 ≈ ~50 µg/mL for dsDNA (literature). These are general QC guidelines, not specifications for this item.

Notes

• Without the exact sequence/length, computed MW, Tm, or extinction coefficients cannot be provided for this specific plasmid. Refer to the CoA/Spec Sheet for itemized values if available.

Quality and Grades

Item-specific facts (from Product Data)

• Grade/purity, QC metrics (e.g., sequence verification, endotoxin level, OD260/280 ratio, restriction digest profile): Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/industry practice)

• Typical QC for CRISPR plasmids may include Sanger sequencing across the sgRNA spacer and scaffold, restriction mapping, agarose gel integrity check (supercoiled fraction), and OD260/280 assessment (~1.8 for dsDNA).
• Application-dependent quality: For mammalian transfection and lentiviral packaging, low endotoxin plasmid prep is preferred to maximize transfection efficiency and cell viability; consider confirming endotoxin level if critical to your workflow.
• Trace components: Buffers often include Tris and EDTA; exact composition and concentrations are item-specific and should be confirmed on the label or CoA.
• Practical implication: If the construct is sequence-verified, maintain the provided lot’s identity by minimizing amplification cycles in bacteria and by re-sequencing the sgRNA region after any recloning step.

Reaction and Applications

Item-specific facts (from Product Data)

• Manufacturer-stated applications: Not specified for this item; refer to catalog/CoA.

General/literature applications for pLenti sgRNA plasmids (context only)

• CRISPR-Cas9 genome editing: The sgRNA cassette directs Cas9 to ABCD3 loci for gene knockout or modulation when co-expressed with Cas9 (via a Cas9 plasmid or Cas9-expressing cell line).
• Lentiviral packaging: The pLenti backbone enables production of replication-incompetent lentiviral particles when co-transfected with packaging plasmids (e.g., psPAX2 and VSV-G) into producer cells (commonly HEK293T). Use BSL-2+ containment and institutional approvals.
• Transient transfection or stable delivery: Transfect directly into cells that already express Cas9 or deliver via lentiviral transduction for stable sgRNA expression.
• Screening: Can be used as a single sgRNA construct in arrayed formats; for pooled screens, verify compatibility with libraries and barcoding.

Practical tips (general)

• Verify spacer sequence integrity by Sanger sequencing before critical experiments.
• Use high-purity, low-endotoxin plasmid DNA for transfection; confirm concentration by A260 and integrity by gel.
• For lentiviral work, optimize transfection reagent, DNA ratios, and harvest times; titer virus and validate on target cells before scale-up.

Reaction Conditions

Item-specific, validated conditions are Not specified for this item; refer to CoA/Spec Sheet and your institutional SOPs.

General/literature guidance for common workflows (not product specs)

• Transient transfection (plasmid DNA): Use high-purity, low-endotoxin DNA. Typical DNA amounts range from 0.5–2 µg per well in 6-well plates; optimize DNA:reagent ratios per transfection reagent instructions. Incubate complexes 10–20 min; change to complete medium after 4–24 h as recommended. Assess sgRNA expression within 24–48 h; edit outcomes often measured 48–96 h post-transfection.
• Lentiviral packaging: Co-transfect sgRNA plasmid with packaging mix (e.g., psPAX2 and pMD2.G/VSV-G) into HEK293T cells at 60–80% confluence. Harvest supernatant at 48 and 72 h, clarify by filtration, and optionally concentrate. Determine functional titer. Perform all steps under BSL-2+ with appropriate PPE and engineering controls.
• Cell transduction: Aim for low MOI (e.g., 0.3) for single-copy integration in pooled screens; include polybrene if compatible. Select with the appropriate antibiotic marker if present (marker not specified for this item).
• Validation: Confirm on-target editing by Sanger/NGS (e.g., TIDE/ICE, amplicon sequencing) and functional assays. Evaluate potential off-targets predicted by design tools.

Safety and Handling

Item-specific facts (from Product Data)

• GHS signal word, hazard statements, pictograms, classification: Not specified for this item; refer to SDS.
• Storage condition: Store at -80°C (provided).
• Shipping: Shipped on dry ice packs + cold packs (provided).

General laboratory safety (literature/guidance; defer to SDS/IBC)

• PPE: Use lab coat, disposable gloves, and eye protection when handling nucleic acids and associated reagents.
• Biosafety: This product is a DNA plasmid in a lentiviral backbone. Plasmid DNA itself is not infectious; however, when used for lentiviral packaging, institutional BSL-2 with enhancements is typically required, along with IBC approval and appropriate training. Follow local biosafety policies.
• Aseptic technique: Work in a clean area with nuclease-free plastics; change gloves frequently to avoid nuclease contamination.
• Incompatibilities: Avoid strong oxidizers, nucleases (RNase/DNase contamination), and repeated freeze–thaw which can shear DNA.
• First aid (general): If exposure to associated buffers/reagents occurs, rinse affected area with water. If eye contact, flush with water for 15 minutes and seek medical advice. In case of ingestion or inhalation of aerosols from associated chemicals, seek medical attention. Always consult the SDS for authoritative guidance.
• Waste: Deactivate biological materials per institutional policy; treat EtBr or other intercalating dyes as hazardous chemical waste.

Solvent Selection

Not typically applicable.

• This product is a plasmid DNA reagent, normally supplied in an aqueous buffer (e.g., nuclease-free water or TE). Organic solvent selection is not relevant. For downstream applications, maintain nuclease-free aqueous conditions and avoid solvents that denature DNA (phenol, chloroform, high % alcohols) unless performing extractions.
• For precipitation or cleanup (general practice), ethanol/isopropanol precipitation and aqueous spin-column elution are standard, but these are process steps rather than solvent choices for use.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage condition: Store at -80°C.
• Shipping: Shipped on dry ice packs + cold packs.

General handling guidance (literature; not product specs)

• Thawing: Thaw on ice. Mix gently by slow pipetting; avoid vigorous vortexing which can shear DNA.
• Aliquoting: Upon first thaw, aliquot into nuclease-free tubes to minimize freeze–thaw cycles. Store working aliquots at -20°C (short term) and archival stocks at -80°C.
• Buffer: If supplied in low-salt buffer (e.g., TE), it may be used directly for cloning or packaging; for transfection, some users dilute in nuclease-free water to minimize EDTA carryover. Exact buffer for this item is Not specified; refer to CoA/Spec Sheet.
• Concentration and purity check: Measure A260/A280 and run an agarose gel to confirm integrity before critical experiments.
• Avoid contamination: Use filtered tips and DNase/RNase-free consumables. Maintain a clean area for plasmid work.

Reconstitution

• If received lyophilized (not indicated for this item), reconstitute in nuclease-free water or TE to the desired concentration, spin briefly to collect, and allow complete dissolution on ice. For this item, reconstitution needs are Not specified; refer to CoA/Spec Sheet.

Structure and Identity

Item-specific facts (from Product Data)

• Product name: pLenti-ABCD3-sgRNA (SKU: P743541)
• Type: Lentiviral CRISPR sgRNA plasmid/vector targeting ABCD3 (inferred from name)
• CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.

General/literature context (for orientation; not item-specific specs)

• Macromolecule class: Circular double-stranded DNA plasmid used to express a single-guide RNA (sgRNA) within a lentiviral backbone for packaging into lentiviral particles or for direct transfection.
• Structural features (2D description): Typical lentiviral sgRNA plasmids include an sgRNA expression cassette (e.g., U6 promoter – sgRNA scaffold with a gene-specific spacer targeting ABCD3), a lentiviral 5’ and 3’ LTR, ψ packaging signal, and common selectable markers and origins of replication; exact map and sequence are item-specific and not provided here.
• Molecular formula / exact molecular weight: Not meaningful for heterogeneous plasmid preparations; precise base pair length and sequence are required and are Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

This section is not typically applicable to plasmid DNA reagents.

• For organic synthesis or small-molecule building-block chemistry, DNA plasmids are not used as reagents. Instead, they are genetic tools for cellular engineering and functional genomics. Relevant details for this product are better found under Reaction & Applications and Reaction Conditions, which cover transfection and lentiviral workflows.

If chemical conjugation to DNA (e.g., click-modified sgRNAs) is of interest, such modifications are outside the scope of this item and would require custom oligonucleotide synthesis.

Target Specificity

Item-specific facts (from Product Data)

• Target: ABCD3 (inferred directly from product name pLenti-ABCD3-sgRNA).
• sgRNA spacer sequence, PAM, target exon/intron, species specificity: Not specified for this item; refer to CoA/Spec Sheet.

General notes (context; not product specs)

• sgRNA activity depends on spacer sequence, genomic context, PAM availability (e.g., NGG for SpCas9), chromatin accessibility, and cell type. Off-target risks increase with near-cognate sites differing by a few mismatches, especially in PAM-proximal regions. Empirical validation is required.