pLenti-ASB2-sgRNA

Cat. No.: P743932

Specifications

Product Name

pLenti-ASB2-sgRNA

Product Line

Biologische Produkte

Lagerung und Versand

Storage

Store at -80°C

Verschickt in

Dry ice packs + Cold packs

Stabilität und Lagerung

Stored at -20 ℃, valid for at least 6 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

Only product-data-sourced protocols and conditions may be listed here.

• Tested applications and recommended conditions: Not specified for this item; refer to CoA/Spec Sheet.

General reminder:

• Follow your institution’s validated protocols for plasmid handling, cloning, transfection, and—if applicable—lentiviral packaging and transduction. This listing does not include item‑specific protocol parameters.

Biological Roles

Item-specific biological role or target is not provided in the Product Data.

General role of sgRNA constructs in CRISPR systems (literature):

• The single guide RNA (sgRNA) forms a ribonucleoprotein complex with a CRISPR-associated protein (e.g., Streptococcus pyogenes Cas9). The 20‑nt spacer within the sgRNA base‑pairs with genomic DNA adjacent to a PAM site, conferring sequence specificity.
• Functional outcomes depend on the CRISPR effector: wild‑type Cas9 introduces double‑strand breaks repaired by NHEJ (yielding indels/knockouts); dCas9‑KRAB represses transcription (CRISPRi); dCas9‑activator fusions recruit transcriptional coactivators (CRISPRa).
• Lentiviral sgRNA vectors enable stable integration or episomal persistence (system‑dependent) for long‑term perturbation and pooled genetic screens.
• Biological considerations: guide efficiency varies with chromatin context, GC content, and sequence features; off‑target potential must be assessed computationally and empirically (e.g., DISCOVER‑Seq, GUIDE‑seq, or targeted amplicon NGS).

Important caveat:

• The specific spacer sequence, PAM context, promoter, and any selectable markers for this SKU are not provided here. Consequently, no claims are made about the biological target or performance of this particular item. Consult the CoA/Spec Sheet for definitive, lot‑specific information.

Buffer Applications

Not typically applicable as a standalone topic. This product is a nucleic acid construct; it is generally stored and handled in simple aqueous buffers (e.g., nuclease‑free water or Tris‑EDTA). For practical handling, see Storage & Reconstitution. For experimental use, buffer systems are dictated by the downstream application (e.g., transfection, viral packaging), which are not specified for this item.

Green Alternatives

While small‑molecule solvent substitution is not relevant here, greener practices can be applied to nucleic acid workflows (general guidance):

• Waste minimization: Use mini‑scale pilot transfections to optimize conditions before scaling viral production, reducing consumables and chemical waste.
• Safer reagents: Prefer non‑toxic transfection agents and aqueous buffers; avoid phenol‑chloroform DNA purification where column- or magnetic bead‑based methods suffice.
• Energy efficiency: Consolidate −80 °C storage to reduce freezer load; aliquot to minimize repeated door openings and freeze–thaw cycles.
• Documentation and reuse: Maintain accurate plasmid maps and sequence files to avoid redundant cloning and repeat orders, saving materials and shipping emissions.

Comparison (general):

• Traditional mutagenesis (chemical/UV) vs. CRISPR plasmid approach: CRISPR/sgRNA systems enable targeted perturbations, decreasing off‑target damage and hazardous chemical usage inherent to random mutagenesis (literature perspective). Trade‑offs include the need for biosafety controls and validation of on‑target edits.

Note: These are general sustainability practices; no item‑specific green certifications or attributes are provided in the Product Data.

Pharmaceutical Uses

No pharmaceutical or clinical use is stated or implied. Research Use Only.

General context (non-clinical):

• Plasmid and lentiviral sgRNA constructs are widely used in discovery research, target validation, and functional genomics screening in preclinical laboratory settings. Any use in manufacturing, diagnostic, or therapeutic contexts would require additional qualification and regulatory oversight not addressed here.

Item-specific regulatory/designation information:

• Pharmacopeial status, GMP grade, or clinical suitability: Not specified for this item; refer to CoA/Spec Sheet.

Users should treat this reagent strictly as a laboratory research tool.

Physical Properties

Item-specific properties:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, buffer, and OD260 purity ratios: Not specified for this item; refer to CoA/Spec Sheet.
• Exact molecular weight, extinction coefficient, and pH: Not specified for this item; refer to CoA/Spec Sheet.

General characteristics of plasmid DNA reagents (literature/general):

• Physical state: Typically supplied as an aqueous solution or lyophilized solid. Solutions often contain Tris‑EDTA or nuclease‑free water; lyophilizates reconstitute in nuclease‑free water or buffer.
• Thermal behavior: DNA does not have a discrete boiling/melting point like small molecules; strand denaturation occurs over a temperature range dependent on GC content and ionic strength.
• Solubility: Highly soluble in aqueous buffers; insoluble in nonpolar organic solvents. Precipitable with alcohols (ethanol/isopropanol) in the presence of salts.
• Stability: Sensitive to nucleases and repetitive freeze–thaw cycles; EDTA and low temperatures improve stability by chelating divalent cations required for nuclease activity.
• Spectroscopy: Quantified by UV absorbance at 260 nm (A260 = 1 corresponds to ~50 µg/mL for dsDNA; literature). A260/A280 ~1.8 indicates low protein contamination; A260/A230 >2.0 suggests low chaotrope/organic carryover.

Note: Since no item-specific measurements are provided in the Product Data, all numerical values above are literature conventions and should not be interpreted as this item’s specifications.

Quality and Grades

Item-specific quality information:

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

How to interpret grades for nucleic acid reagents (general guidance):

• Research-use plasmid DNA is commonly qualified by OD260/280 and OD260/230 ratios, supercoiled fraction, and absence of endotoxin if intended for cell transfection. If endotoxin-reduced or transfection-grade is required, this is typically stated explicitly on the CoA. No such designation is provided here.
• Sequence identity and insert verification are ordinarily supported by restriction analysis and/or Sanger/NGS readout. For sgRNA vectors, guide spacer sequence and any cloning scar details are essential for experimental reproducibility.
• Sterility is typically “not sterile” for plasmid preparations unless indicated. If sterility or mycoplasma-tested status is needed for sensitive cell systems, confirm on the CoA.

What is specific to this item:

• The Product Data do not provide sequence map, spacer, antibiotic marker, promoter identity, or endotoxin level. Users should obtain the CoA/Spec Sheet for authoritative specifications before critical experiments.

Recommendations:

• If using for viral packaging or sensitive primary cells, confirm endotoxin level and supercoiled fraction on the CoA.
• Archive documentation (lot-specific CoA and sequence files) with your ELN/LIMS to ensure traceability.

Reaction and Applications

Item-specific applications provided: None listed in the Product Data.

General applications for sgRNA lentiviral plasmids (literature/typical workflows; not item-specific):

• CRISPR gene perturbation: Express an sgRNA to guide a CRISPR effector (e.g., Cas9 for nuclease activity, dCas9‑KRAB for CRISPRi, dCas9‑VPR for CRISPRa) to a genomic locus. The precise target sequence for this SKU is not specified here.
• Stable delivery: When co-transfected with compatible packaging plasmids, pLenti‑backbone sgRNA vectors enable production of pseudotyped lentiviral particles for efficient transduction of dividing and non‑dividing mammalian cells, allowing pooled or arrayed screens.
• Library applications: SgRNA vectors are used to construct pooled libraries for CRISPR knockout/CRISPRi/CRISPRa screening. Key parameters include MOI control, coverage, and representation.
• Cloning and validation: Typical steps include oligo design (PAM‑adjacent 20 nt spacer), annealing/ligation or Golden Gate cloning into the sgRNA scaffold, colony PCR, and Sanger validation. U6/H1 promoters require accurate transcription start and no 5′-G loss; spacer design may include a leading “G” if needed.

Practical tips (general):

• Maintain nuclease‑free technique and verify insert sequence before scale‑up. For viral work, follow BSL‑2 practices and include proper controls (non‑targeting sgRNA, positive controls). Optimize antibiotic selection and timing if the vector includes a selection cassette. The exact selectable marker and promoter(s) for this item are not provided—consult the CoA.

Reaction Conditions

No item-specific reaction conditions are provided in the Product Data.

General laboratory conditions (literature; not item-specific):

• Cloning: Golden Gate assembly with type IIS enzymes (e.g., BsaI/BsmBI) is commonly performed at 37 °C with cycling to 16 °C, followed by a 50–65 °C heat-inactivation step. Ligation reactions are typically incubated 5–60 min depending on protocol and enzyme system.
• Transformation: Chemically competent E. coli (e.g., 10^6–10^9 cfu/µg) are heat‑shocked at 42 °C for ~30–60 s; electrocompetent cells are pulsed at 1.8 kV (0.1 cm cuvette) followed by recovery at 30–37 °C for 30–60 min in SOC.
• Viral packaging (if applicable to the user’s workflow): HEK293T cells are commonly transfected with transfer, packaging, and envelope plasmids; supernatant harvest at 48–72 h post‑transfection. Biosafety and precise plasmid ratios are lab‑specific and not provided here.

These are general practices and do not constitute specifications or validated protocols for this item. For lot‑specific instructions, consult the CoA/Spec Sheet and your institutional SOPs.

Safety and Handling

Item-specific hazard information from Product Data:

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

General safety guidance for nucleic acid research reagents (literature/best practice):

• Handle using good microbiological practice in a clean area dedicated to molecular biology. Wear appropriate PPE: lab coat, gloves (nitrile), and safety glasses.
• Avoid nuclease contamination: use certified nuclease‑free consumables and change gloves frequently; keep containers closed to minimize airborne nuclease exposure.
• If the plasmid is used to generate lentiviral particles, follow institutional biosafety procedures for viral vector work (e.g., BSL‑2 practices, biological safety cabinet, appropriate waste deactivation). This product listing does not specify packaging status; consult the SDS and your IBC for requirements.
• Incompatibilities: Avoid strong nucleases (DNase), prolonged exposure to elevated temperature, and repeated freeze–thaw. Keep away from oxidizing decontaminants that can damage DNA (e.g., hypochlorite) unless deactivation/disposal is intended.
• First aid (overview; defer to SDS): After skin/eye contact with solutions or buffers, rinse with water. If inhaled aerosols or ingestion occur, seek medical attention following institutional protocols.

Always consult the product-specific SDS and institutional biosafety policies before use.

Solvent Selection

Not typically applicable. This product is a nucleic acid research reagent rather than a small-molecule solute. It is generally supplied in, and used with, aqueous buffers that maintain DNA integrity (e.g., nuclease‑free water, Tris‑EDTA). For handling considerations, see Safety & Handling and Storage & Reconstitution.

Storage and Reconstitution

Item-specific storage and shipping (from Product Data):

• Storage: Store at −80 °C.
• Shipped in: Dry ice packs + Cold packs.

Additional handling guidance (general best practices; not item-specific specifications):

• Upon receipt: Verify package integrity and store immediately at −80 °C. If provided lyophilized, allow to equilibrate to room temperature in a desiccated environment before opening to prevent condensation.
• Aliquoting: To minimize freeze–thaw cycles, prepare single‑use aliquots in nuclease‑free tubes. Avoid repeated freeze–thaw as it can degrade DNA and reduce performance.
• Reconstitution (if lyophilized): Use nuclease‑free water or low‑salt buffer (e.g., 10 mM Tris‑HCl, 0.1–1 mM EDTA, pH ~8.0). Mix gently by pipetting; avoid vigorous vortexing that can shear DNA.
• Thawing: Thaw solutions on ice, mix gently, and briefly spin down to collect contents. Keep on ice during setup; return unused aliquots promptly to −80 °C.
• Documentation: Record lot number, concentration, and buffer composition from the CoA/Spec Sheet in your ELN/LIMS. Maintain a working map and verified sequence for traceability.

Note: Where exact buffer composition, concentration, or stabilizers are required, consult the lot‑specific CoA/Spec Sheet. Research Use Only.

Structure and Identity

• SKU: P743932
• Product name: pLenti-ASB2-sgRNA

Item-specific identifiers (from Product Data):

• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• CID: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature/typical for naming):

• The prefix “pLenti” commonly denotes a plasmid backbone compatible with lentiviral production workflows (third‑generation systems are typical in research). The suffix “sgRNA” indicates the presence of a single guide RNA expression cassette used for CRISPR applications (e.g., Cas9 targeting).
• Such vectors generally comprise: a bacterial origin of replication and antibiotic resistance marker for propagation in E. coli; a U6/H1 (or similar) RNA polymerase III promoter driving sgRNA; and lentiviral long terminal repeat (LTR) and packaging signals for viral particle production when co-transfected with helper plasmids. Exact features for this item are not specified.

2D/functional features (general, not item-specific):

• Biomacromolecule category: circular double‑stranded DNA plasmid (if supplied as plasmid) or nucleic acid cassette; not a small molecule.
• Functional elements typically include: promoter(s), sgRNA scaffold, cloning site for spacer/target sequence, and selection markers. Specific map, insert, and sequence for this SKU are not provided here; consult the CoA/Spec Sheet for definitive details.

Synthetic Utility

Not applicable in the classical organic synthesis sense. This product is a nucleic acid research reagent rather than a small‑molecule building block.

General note (for molecular biology workflows):

• As a plasmid/backbone for sgRNA expression, it serves as a genetic “scaffold” into which variable spacers can be cloned for targeted genome engineering. Utility lies in modular cloning (e.g., Golden Gate with type IIS restriction enzymes) rather than chemical transformations.

For actionable details on cloning strategies, refer to Reaction & Applications and consult the item’s CoA/Spec Sheet for any vector‑specific requirements.

Target Specificity

Only product-data-sourced information may be listed here.

• Target gene/locus: Not specified for this item; refer to CoA/Spec Sheet.
• Species reactivity or cell system: Not specified for this item; refer to CoA/Spec Sheet.
• Vector map, promoter(s), selection marker(s), and sgRNA spacer sequence: Not specified for this item; refer to CoA/Spec Sheet.

Note: Although the product name suggests an sgRNA context, no definitive target details are provided in the Product Data for this SKU.