pLenti-OLIG2-sgRNA

Cat. No.: P748713

Specifications

Product Name

pLenti-OLIG2-sgRNA

Product Line

biological_products

Lagerung und Versand

Storage

Store at -80°C

Verschickt in

Dry ice packs + Cold packs

Stabilität und Lagerung

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

Application Protocols

Only provide information from product data.

• Tested/validated applications (e.g., WB, IHC, IF, FC for antibodies; transfection/transduction parameters for plasmids), recommended dilutions/MOIs, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.

General guidance: See Reaction Conditions for literature-based workflows that may serve as a starting point; optimize empirically for your system.

Biological Roles

Item-specific biological validation: Not specified for this item; refer to CoA/Spec Sheet.

Gene target context (literature/general):

• OLIG2 (oligodendrocyte lineage transcription factor 2) is a basic helix-loop-helix (bHLH) transcription factor central to oligodendrocyte and certain motor neuron lineage specification. It regulates progenitor proliferation and differentiation programs in the central nervous system.
• In developmental neurobiology, OLIG2 is implicated in ventral spinal cord patterning and myelination pathways. Alterations in OLIG2 expression impact glial fate decisions.

Relevance to sgRNA-based studies (general):

• CRISPR-mediated disruption of OLIG2 can be used to probe its role in oligodendrocyte maturation, interplay with SOX10 and NKX2.2, and downstream myelin gene expression. In stem-cell-derived neural models, stable sgRNA expression via lentiviral vectors allows longitudinal studies of lineage commitment (research use only).
• Complementary assays include qPCR for lineage markers (e.g., MBP, CNP), immunocytochemistry for OLIG2/SOX10, and functional readouts like myelination co-culture assays (application-dependent).

Note: The precise guide sequence(s), species targeting (human/mouse/rat), and any validation data are Not specified for this item; consult the CoA/Spec Sheet for construct-specific details.

Buffer Applications

This product is a plasmid/sgRNA construct rather than a buffering agent. Traditional buffer system selection (pH range, capacity) is not directly applicable.

Practical notes (general):

• Storage/handling buffers: Nuclease-free water or TE buffer (10 mM Tris-Cl, 0.1–1 mM EDTA, pH 7.5–8.5) are commonly used to preserve plasmid integrity and reduce nuclease activity.
• Enzymatic reactions: Downstream cloning, restriction digests, and PCR require specific manufacturer-provided reaction buffers; select buffers compatible with the enzymes in use.

Item-specific buffer composition: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Green chemistry framing is not directly applicable to a plasmid DNA reagent. Nevertheless, the upstream and downstream workflows can be made greener (general guidance):

• DNA prep and purification:
  • Prefer column-based or magnetic-bead purification kits that avoid large volumes of phenol/chloroform. If organic extraction is required, implement microscale procedures and proper waste segregation.
  • Use ethanol/isopropanol precipitation with minimal solvent volumes and recover/neutralize alcohol waste per institutional guidelines.
• Viral packaging and cell culture:
  • Minimize single-use plastics by adopting reusable serological pipettes where validated and employing bulk media preparation to reduce packaging waste.
  • Optimize transfection conditions to lower DNA and reagent usage while maintaining titer.
• Energy usage:
  • Consolidate freezer inventory and employ high-efficiency -80°C freezers; use set-point warm-up strategies during defrost cycles to reduce energy consumption without compromising sample integrity.

Illustrative comparison (general):

• Traditional phenol/chloroform plasmid purification: High hazard, significant solvent waste, strong PPE needs.
• Column/bead-based plasmid purification: Aqueous buffers, reduced hazard, lower VOC emissions, often comparable yield and purity.

For this specific item, no manufacturing green metrics are specified; refer to CoA/Spec Sheet for any sustainability notes.

Pharmaceutical Uses

This reagent is designated For research use only and is not intended for human or veterinary use, diagnostic procedures, or as a therapeutic.

Formulation/excipient context: Not applicable. Plasmid/viral vector reagents in a research setting are used for in vitro and in vivo laboratory studies and are not pharmaceutical-grade materials unless explicitly stated.

Regulatory notes (general):

• RUO materials are not manufactured under GMP for clinical applications. Any translation to regulated use would require re-sourcing or requalification under appropriate quality systems.

Item-specific pharmacopeial/compendial status: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific physico-chemical data: Not specified for this item; refer to CoA/Spec Sheet.

General properties for plasmid DNA solutions (literature/general):

• Physical state: Aqueous solution or hydrated film/pellet of circular dsDNA; often supplied in TE buffer or water. Actual appearance/buffer for this item: Not specified.
• Solubility: Plasmid DNA is water-soluble; insoluble in most organic solvents (ethanol/isopropanol precipitate DNA rather than dissolve it).
• UV absorbance: A260 used for quantification (1 A260 ≈ 50 µg/mL for dsDNA; literature value). A260/A280 ~1.8 indicates low protein contamination; A260/A230 >2.0 indicates low salt/phenol carryover.
• pKa/logP: Not applicable to high-molecular-weight polyanions like DNA; characterization is via biophysical measurements rather than small-molecule descriptors.
• Melting/boiling point: Not applicable (biopolymer). Thermal denaturation (Tm) is sequence/length dependent and not a fixed value.
• Density/refractive index of buffer: Determined by the aqueous buffer composition; Not specified for this item.

Concentration and form at shipment/use: Not specified for this item; refer to CoA/Spec Sheet. If received dried, rehydrate with nuclease-free water or buffer (see Storage & Reconstitution).

Quality and Grades

Item-specific grade/purity and QC attributes: Not specified for this item; refer to CoA/Spec Sheet.

Context for plasmid/sgRNA reagent quality (general/literature):

• Sequence identity: High-quality plasmids for CRISPR applications are typically sequence-verified across the sgRNA insert and essential backbone features. Verification method might include Sanger sequencing using U6 and scaffold primers. Status for this item: Not specified.
• Endotoxin level: For cell culture and transduction, low-endotoxin DNA (<10 EU/µg, literature) is often preferred to minimize cytotoxicity. Status for this item: Not specified.
• Topology: Supercoiled fraction supports efficient transfection; nicked/linear content can reduce performance. Status for this item: Not specified.
• Sterility and nuclease-free handling: Critical to prevent degradation/contamination. Status for this item: Not specified.
• Packaging: DNA is often supplied in nuclease-free water or TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0). Exact buffer for this item: Not specified.

How to interpret “research use only”:

• RUO materials are intended solely for laboratory research. They are not validated for diagnostic, clinical, or therapeutic use.

Recommendation: Consult the CoA/Spec Sheet for plasmid map, sequence of the OLIG2 guide(s), selection marker(s), antibiotic resistance, recommended growth strain, and any verified QC metrics specific to SKU P748713.

Reaction and Applications

Item-specific validated applications: Not specified for this item; refer to CoA/Spec Sheet.

General/literature applications for pLenti sgRNA reagents targeting transcription factors like OLIG2:

• CRISPR-Cas9 gene editing: The sgRNA provides sequence-specific guidance to Cas9 (e.g., SpCas9) to introduce double-strand breaks at OLIG2 loci, enabling knockout via NHEJ indels or precise edits via HDR when a donor template is provided.
• Stable delivery via lentiviral transduction: Lentiviral backbones facilitate integration into dividing and non-dividing mammalian cells for durable sgRNA expression, useful in long-term differentiation studies of glial lineages.
• Functional genomics: Use in loss-of-function screens to interrogate OLIG2 roles in lineage specification, myelination pathways, or neural progenitor biology (research use only).
• Reporter/selection coupling: If the vector contains a selection marker (e.g., puromycin), post-transduction selection enriches sgRNA-positive cells. Marker identity for this item: Not specified.

Practical tips (general):

• Guide design: Validate PAM-compatible, off-target-minimized guide sequences against the relevant genome build. For multiple guides, consider tiling across early exons.
• Controls: Include non-targeting sgRNA and positive control sgRNA (targeting an essential gene) to benchmark editing efficiency.
• Verification: Assess editing via T7E1 or TIDE/ICE analysis, targeted amplicon NGS, and confirm functional knockout at mRNA/protein levels.
• Packaging: Co-transfect with packaging (e.g., psPAX2) and envelope (e.g., VSV-G) plasmids; titrate viral supernatant, and transduce at defined MOIs.

Reaction Conditions

Small-molecule reaction conditions are not applicable. For molecular biology workflows involving pLenti sgRNA constructs (general/literature guidance):

• Transfection for lentiviral packaging: HEK293T cells at 60–80% confluency; co-transfect sgRNA vector with packaging plasmids using lipid-based transfection reagents in serum-free medium, then replace with complete medium after 6–12 h. Harvest viral supernatant at 48–72 h. Optimize DNA:reagent ratios per vendor recommendations.
• Transduction: Add filtered viral supernatant with polybrene (e.g., 4–8 µg/mL; verify cell tolerance) to target cells. Spinoculation may improve MOI. Replace media after 12–24 h. Select with the appropriate antibiotic if the vector encodes resistance (marker Not specified for this item).
• Editing window: Assess genomic modification 48–96 h post-transduction for initial indels; clonal isolation may be required for complete knockout.
• Controls and readouts: Include non-targeting and positive controls; quantify editing via Sanger/NGS of amplicons and evaluate OLIG2 expression by qPCR/Western/ICC.

All times, concentrations, and additives above are general literature practices and should be optimized per cell type. Product-specific protocols are Not specified for this item.

Safety and Handling

GHS/SDS fields for this specific item:

• Signal word, H-statements, GHS classification, pictograms: Not specified for this item; refer to SDS.

General laboratory safety guidance for plasmid/lentiviral sgRNA constructs (literature/best practice):

• Biosafety: Handle plasmid DNA using standard microbiological practices. If producing lentiviral particles using this construct, work under BSL-2 containment (or as required by your IBC) with appropriate engineering controls (biosafety cabinet, sealed rotors), since replication-incompetent lentiviral vectors are risk group 2.
• PPE: Lab coat, gloves, eye protection. Change gloves frequently when handling DNA to avoid nuclease contamination.
• Incompatibilities: Avoid contaminating DNA with nucleases (RNase for sgRNA in vitro transcription, DNase for plasmid), strong oxidants, and non-sterile solutions. Do not mix with phenol/chloroform outside a fume hood.
• First aid (general): In case of contact with eyes/skin, rinse with water. If inhaled (aerosols from lyophilized material) move to fresh air. If ingested, rinse mouth. Seek medical attention as per institutional guidelines and SDS.
• Waste: Deactivate residual lentiviral particles with appropriate disinfectants (e.g., 10% bleach, 70% ethanol contact times per policy). Autoclave biological waste.
• Transport: Keep cold to preserve nucleic acid integrity; avoid repeated freeze–thaw cycles.

Always defer to the product-specific SDS and your institutional biosafety committee for authoritative safety requirements.

Solvent Selection

This product is a nucleic acid construct, not a small-molecule solute. Classical solvent selection frameworks (polarity/logP) are not directly applicable.

Practical handling media (general guidance):

• Aqueous buffers: Nuclease-free water or low-salt buffers (e.g., TE: 10 mM Tris-Cl, 0.1–1 mM EDTA, pH 7.5–8.5) are commonly used to maintain plasmid stability and chelate divalent cations that promote nuclease activity.
• Cell transfection/transduction: Mix with transfection reagents (lipid/polymer-based) in serum-free media per reagent instructions. For lentiviral packaging, DNA is combined with packaging/helper plasmids and transfected into producer cells (e.g., HEK293T) using suitable transfection buffers.
• Precipitation: Ethanol/isopropanol precipitate DNA for concentration or desalting; avoid overdrying the DNA pellet.

When to choose alternative media:

• For immediate cloning or in vitro transcription of sgRNA, nuclease-free water may be preferred to minimize EDTA carryover.
• For long-term plasmid storage, TE provides improved stability versus water at 4°C or room temperature; however, for -80°C storage, water or TE are both commonly used.

Note: Organic solvents used in small-molecule workflows are not pertinent here; maintain aqueous, sterile, nuclease-free conditions.

Storage and Reconstitution

Item-specific conditions from Product Data:

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

Best practices (general) for plasmid/sgRNA constructs:

• Upon receipt: Verify package integrity; keep frozen at -80°C until use. Minimize time at ambient temperature during unpacking.
• Thawing: Thaw on ice. Briefly spin down contents to collect condensate. Keep on ice during handling to reduce nuclease activity.
• Aliquoting: Prepare single-use aliquots in sterile, nuclease-free tubes to avoid repeated freeze–thaw cycles (which can nick DNA and reduce transfection efficiency).
• Storage buffers: Nuclease-free water or TE buffer (10 mM Tris-Cl, 0.1–1 mM EDTA, pH 7.5–8.5) are commonly used. If supplied dry, reconstitute with nuclease-free water or TE to a convenient working concentration (item-specific concentration/volume: Not specified for this item; refer to CoA/Spec Sheet).
• Short-term storage: 4°C for up to a week in TE is common; for longer-term, -20°C or -80°C is preferred. Avoid multiple freeze–thaw cycles.
• Documentation: Record lot number, concentration (A260), buffer composition, and date of first thaw.

Always refer to the product’s CoA/Spec Sheet for any construct-specific reconstitution instructions or stabilizers not listed here.

Structure and Identity

Item-specific identifiers (CAS, CID, InChIKey, SMILES, exact sequence/backbone map): Not specified for this item; refer to CoA/Spec Sheet.

What the product name implies (general/literature context, not item-specific specs):

• pLenti-OLIG2-sgRNA denotes a lentiviral plasmid or lentiviral-ready construct designed to express a single-guide RNA (sgRNA) targeting the OLIG2 gene (oligodendrocyte lineage transcription factor 2).
• Typical architectural features of pLenti sgRNA vectors (literature/general):
  • A lentiviral backbone with 5′ and 3′ LTRs and ψ packaging signal to enable production of pseudotyped lentiviral particles in trans.
  • An RNA polymerase III promoter (commonly U6 or H1) driving the sgRNA scaffold containing a 20-nt guide sequence (spacer) specific to OLIG2.
  • Optional selection markers (e.g., puromycin, blasticidin) and/or fluorescent reporters for selection/tracking—presence and identity Not specified for this item; refer to CoA/Spec Sheet.
  • For CRISPR workflows, the Cas9 nuclease is typically supplied on a separate plasmid unless the vector is “all-in-one”; configuration Not specified for this item.

2D/functional description (general):

• The functional “structure” is a DNA plasmid in a circular, double-stranded form. The sgRNA expression cassette produces a short noncoding RNA that complexes with Cas9 to direct sequence-specific double-strand breaks at OLIG2 genomic loci harboring a compatible PAM (e.g., NGG for SpCas9).

Molecular formula and molecular weight: Not specified for this item; refer to CoA/Spec Sheet (depends on exact backbone length and inserts).

Synthetic Utility

Classical small-molecule synthetic utility is not applicable to a plasmid DNA construct. However, from a molecular biology “construct engineering” perspective (general):

• Modular cloning: The sgRNA spacer sequence can often be inserted via annealed oligos into a vector backbone using type IIS restriction enzymes (e.g., BsmBI/BbsI) or Gibson/Golden Gate assembly. This enables rapid re-targeting to different OLIG2 sites or alternative genes.
• Vector customization: Backbones can be re-engineered to swap promoters, selection markers, or to convert sgRNA-only vectors into all-in-one Cas9+sgRNA systems, depending on experimental needs. Specific features for this item are Not specified.
• Functional derivatives: The sgRNA cassette can be adapted for CRISPRi (dCas9-KRAB) or CRISPRa (dCas9-VP64) by using appropriate effector systems to repress or activate OLIG2 expression without cutting.

Note: For precise cloning sites, antibiotic resistance markers, and promoter identities for SKU P748713, refer to the product’s plasmid map/sequence (Not specified here).

Target Specificity

Only provide information from product data.

• Target/antigen/gene details, species reactivity, clone/isotype (if antibody), epitope/guide sequence(s), and validation: Not specified for this item; refer to CoA/Spec Sheet.

Note: Although the product name implies sgRNA targeting OLIG2, the exact guide sequence(s), PAM context, and species specificity are not provided here.