pLenti-YTHDC2-sgRNA

Cat. No.: P751893

Especificaciones

Product Name

pLenti-YTHDC2-sgRNA

Product Line

Productos biológicos

Almacenamiento y envío

Condiciones de almacenamiento de almacenamiento

Store at -80°C

Enviado en

Dry ice packs + Cold packs

Estabilidad y almacenamiento

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

Application Protocols

Per instruction, this section may only include protocols explicitly provided in the Product Data.

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

For general workflow guidance, see the Reaction Conditions tab.

Biological Roles

Focus gene: YTHDC2 (as indicated by product name; specific species/guide not provided).

Literature/general overview of YTHDC2 biology:

• Function: YTHDC2 is an RNA helicase and m6A reader protein implicated in the regulation of RNA stability and translation. It contains a YTH domain that recognizes N6‑methyladenosine (m6A) marks on RNA and helicase motifs that remodel RNA–protein complexes.
• Cellular processes: Reported roles include modulation of mRNA turnover, translation efficiency, and regulation of meiotic progression in germ cells. YTHDC2 participates in RNA surveillance pathways and interacts with decay machinery.
• Pathways: Interfaces with m6A-dependent post‑transcriptional regulation; may coordinate ribonucleoprotein assembly and 3′‑UTR dynamics.
• Research utility of sgRNA targeting YTHDC2: Enables perturbation of YTHDC2 to study m6A‑dependent RNA metabolism, meiosis‑related programs in relevant models, and synthetic interactions with other RNA-binding proteins.

Caveats:

• Phenotypic outcomes are cell‑type specific and depend on the extent and nature of perturbation (knockout vs. knockdown vs. CRISPRi/a). Use multiple guides and appropriate controls to deconvolute on‑target effects.

Item-specific organism, transcript isoform, exon targeting, and guide sequence are not specified for this SKU; consult the CoA/Spec Sheet for targeting details.

Buffer Applications

Direct use as a buffer is not applicable; this product is a plasmid DNA reagent.

General handling buffers for plasmid DNA (literature/practice):

• Storage buffers: TE (10 mM Tris–HCl, 1 mM EDTA, pH 7.5–8.0) supports long‑term stability by chelating Mg2+ (inhibiting DNases) and maintaining pH.
• Working buffers: Nuclease‑free water or low‑salt Tris for downstream enzymology; PBS or culture media for immediate complexation with transfection reagents.
• Electrophoresis: Agarose gel analysis commonly performed in TAE or TBE; supercoiled vs. nicked forms resolve differently.

Item-specific buffer composition and concentration are not provided; refer to CoA/Spec Sheet.

Green Alternatives

Context: This is a plasmid DNA research reagent. Green chemistry considerations pertain primarily to upstream/downstream workflows (prep, transfection, viral packaging) rather than solvent choice for the item itself.

Opportunities to reduce environmental footprint (literature/practice):

• Minimize solvent use in plasmid prep: Prefer silica spin columns or aqueous precipitation workflows over large‑volume phenol–chloroform extraction. If organic extraction is necessary, ensure proper waste segregation and recovery.
• Low‑endotoxin aqueous systems: Use water-based buffers and avoid halogenated solvents entirely in cell culture and viral packaging.
• Scalable transfection: Use efficient, low‑toxicity, water‑compatible transfection reagents to reduce reagent volumes and cell waste; optimize DNA:reagent ratios to minimize excess.
• Energy and cold chain: Consolidate shipments and store working aliquots at −20°C when feasible for short-term to reduce door‑opening of −80°C freezers; for long-term, follow item storage specs.
• Waste management: Treat lentiviral waste with bleach prior to disposal; neutralize bleach before drain disposal where required to reduce chloride burden; adhere to local regulations.

Trade‑offs:

• Phenol/chloroform delivers high purity but creates hazardous waste; column methods reduce hazard but add solid waste. Evaluate data needs against environmental impact.

Item-specific greener alternatives are not applicable; the product itself is an aqueous DNA reagent.

Pharmaceutical Uses

This product is designated “For research use only” and is not intended for diagnostic, therapeutic, or clinical applications.

Context (general/industry):

• Plasmid DNA reagents like pLenti‑format sgRNA vectors are used in discovery research, target validation, and process development in non‑GMP laboratories. They may inform upstream concepts for gene therapy, but this SKU is not produced to pharmacopeial or GMP specifications.
• Formulation roles: Not applicable as a pharmaceutical excipient. When used in research, plasmids are formulated in aqueous buffers optimized for stability and transfection, not for human administration.

Compliance note: Any translation of research findings toward clinical use requires separate sourcing/manufacture under appropriate GMP and regulatory frameworks. No such claims are made or implied for this item.

Physical Properties

Item-specific physicochemical specifications (concentration, buffer, A260/A280, endotoxin, size in bp): Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties for plasmid DNA vectors:

• Physical state: circular double‑stranded DNA in aqueous buffer or lyophilized (varies by lot; item-specific status not specified).
• Solubility (literature): Highly soluble in aqueous buffers (e.g., TE, Tris, PBS) and water; insoluble in organic solvents such as alcohols, ethers, chlorinated solvents.
• Stability (literature): Stable for months at −20 to −80°C; repeated freeze–thaw can induce nicking and reduced transfection efficiency.
• UV characteristics (literature): DNA λmax ~260 nm; typical A260/A280 of 1.8–2.0 for pure DNA; A260/A230 >2 desirable; these are quality indicators, not product specifications.
• Viscosity/handling: Concentrated plasmid preparations can be moderately viscous; pipet slowly to avoid shearing.
• pH sensitivity: Optimal stability in slightly basic buffers (e.g., 10 mM Tris, pH 7.5–8.5). Acidic conditions promote depurination.
• Thermal behavior: No meaningful boiling/melting point in the classical sense; plasmid secondary structure denatures above ~90–95°C, with renaturation on cooling depending on ionic strength.

Note: Do not interpret the above as item specifications. For exact length (kb), formulation, and concentration of SKU P751893, consult the CoA/Spec Sheet.

Quality and Grades

Item-specific grade/purity, sequencing verification, endotoxin level, antibiotic marker(s), and backbone map: Not specified for this item; refer to CoA/Spec Sheet.

General guidance on quality attributes for plasmid DNA reagents:

• Plasmid QC practices (literature/industry):
  • Identity: Restriction digest pattern and/or full plasmid sequencing to confirm the sgRNA cassette and backbone integrity.
  • Purity: A260/A280 ~1.8–2.0; minimal contaminating RNA/protein. Supercoiled fraction typically preferred for transfection.
  • Endotoxin: Low-endotoxin or endotoxin-free prep recommended for mammalian cell transfection and viral packaging (commonly <0.1 EU/µg for high-sensitivity lines; item-specific value not specified).
  • Sterility: 0.22 µm filtration and bioburden control for cell culture applications.
• Grade definitions (explanatory):
  • Research grade: Suitable for standard cloning, transfection, and virus production in research settings; not for diagnostic or therapeutic use.
  • Transfection grade: Emphasis on low endotoxin and high supercoiled content for improved transfection efficiency.

What is specific for this SKU: Only storage and shipping are specified in the Product Data. For map, selectable marker, and QC metrics, consult the CoA/Spec Sheet.

Reaction and Applications

Manufacturer Applications: Not specified beyond the name; this is a lentiviral sgRNA plasmid intended for research use.

Expanded use cases (literature/general for pLenti sgRNA vectors):

• CRISPR gene knockout/perturbation: Expression of an sgRNA targeting YTHDC2 for use with Cas9-expressing cells or co-delivery of Cas9 (knockout), or with dCas9 effectors for CRISPRi/CRISPRa screens.
• Stable delivery via lentivirus: Packaging the plasmid with helper constructs (gag/pol, rev, env) enables transduction of dividing and non‑dividing cells, integration into host genome, and stable sgRNA expression.
• Functional genomics: Single‑gene validation, construction of isogenic knockouts, or inclusion in pooled libraries centered on RNA metabolism pathways involving YTHDC2.
• Rescue/epistasis studies: Pair with YTHDC2 cDNA rescue or helicase‑dead mutants to dissect on‑target phenotypes.

Practical notes (general):

• Guide performance: Efficacy depends on guide sequence, GC content, PAM compatibility, chromatin context, and cell line. Use multiple independent guides and appropriate controls (non‑targeting sgRNA).
• Validation: Confirm editing/perturbation with TIDE/ICE, amplicon NGS, and protein‑level assays (e.g., Western blot) where applicable.
• Off‑target mitigation: Employ high‑fidelity Cas9 variants, truncated guides, or orthogonal PAMs; verify via targeted off‑target sequencing when critical.

Item-specific guide sequence and backbone features are not provided; consult the CoA/Spec Sheet.

Reaction Conditions

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

General/literature guidance for common workflows with lentiviral sgRNA plasmids:

• Cloning of guide: Anneal and ligate oligos into the sgRNA cassette using vendor‑specified overhangs; transform into high‑efficiency E. coli; confirm by Sanger sequencing.
• Transfection for viral packaging: Co‑transfect HEK293T/293FT cells with pLenti‑sgRNA plasmid plus packaging plasmids (e.g., gag/pol, rev, VSV‑G). Use serum‑compatible transfection reagents; optimize DNA:reagent ratio per manufacturer. Replace medium after 6–24 h.
• Virus harvest: Collect supernatants at 48–72 h; clarify and optionally concentrate (e.g., PEG or ultracentrifugation). Titer by p24 or functional MOI.
• Transduction: Add virus to target cells with polybrene (e.g., 4–8 µg/mL, cell‑line dependent) or spinoculate; select with appropriate antibiotic if the vector encodes a resistance marker. Maintain BSL‑2 procedures.
• Editing assessment: For Cas9‑expressing cells, assess indels 48–96 h post‑transduction; for stable lines, allow sufficient time for turnover of existing protein.

Notes:

• Temperature: Standard mammalian culture conditions (37°C, 5% CO2).
• Controls: Non‑targeting sgRNA, safe‑targeting sgRNA, and multiple independent YTHDC2 guides to confirm on‑target effects.

Adapt specifics (cell line, MOI, selection) to your system.

Safety and Handling

GHS classification, signal word, hazard statements, pictograms: Not specified for this item; refer to SDS.

General biosafety guidance (literature/practice for lentiviral sgRNA plasmids):

• Risk profile: Plasmid DNA itself is generally considered low hazard; however, when used to generate lentiviral particles, handle under BSL‑2 containment following institutional biosafety approvals for recombinant DNA and lentiviral work.
• PPE: Lab coat, nitrile gloves, and eye protection. Use biological safety cabinet (Class II) for any viral packaging or handling of virus‑containing material.
• Incompatibilities: Avoid strong acids/bases that can hydrolyze DNA; avoid nucleases. Maintain nuclease‑free technique (filtered tips, dedicated solutions).
• First aid overview: If skin or eye contact with buffer occurs, rinse with water. If aerosol exposure occurs during viral work, follow spill/exposure response per institutional biosafety procedures and seek medical attention per SDS guidance.
• Decontamination: Surfaces and waste potentially contaminated with lentivirus should be treated with freshly prepared 10% bleach (≥10 min contact) followed by 70% ethanol. DNA solutions without viral content can be disposed per local rules for non-hazardous biotech waste.
• Transport/packaging: Keep cold; protect from nuclease contamination and repeated freeze–thaw cycles.

Always defer to the product’s SDS and your IBC-approved SOPs for authoritative safety requirements.

Solvent Selection

Applicability: This product is a plasmid DNA reagent, not a small‑molecule solute. Traditional organic solvent selection does not apply.

General handling (literature/practice):

• Preferred medium: Nuclease‑free water or aqueous buffers (e.g., TE: 10 mM Tris–HCl, 1 mM EDTA, pH 7.5–8.0) to stabilize DNA against nucleases and depurination. For transfection/packaging, many users dilute in PBS or Opti‑MEM immediately prior to complexation.
• Avoid: Neat organic solvents (ethanol, isopropanol, acetone, chlorinated solvents) which precipitate or denature DNA. If precipitation is desired (e.g., cleanup), use ethanol/isopropanol protocols with appropriate salts, then redissolve in nuclease‑free water/TE.
• Additives: EDTA (0.1–1 mM) chelates Mg2+ to inhibit DNases; avoid EDTA if downstream enzymatic reactions require divalent cations.
• Filtration: Use 0.22 µm sterile, low‑protein‑binding filters for sterile formulations.

Selection tips:

• Long‑term storage: TE at pH ~8 is commonly chosen for plasmid stability (literature).
• Immediate cell culture use: Water or low‑salt buffers minimize interference with transfection reagents; follow reagent vendor guidance.

Storage and Reconstitution

Item-specific conditions from Product Data:

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

Additional handling guidance (general/practice for plasmid DNA; not product specifications):

• Upon receipt: Verify package integrity; minimize thaw time. If supplied frozen, keep on dry ice/ice during handling. Centrifuge briefly to collect contents.
• Aliquoting: Prepare single‑use aliquots in nuclease‑free tubes to avoid repeated freeze–thaw cycles, which can nick DNA and reduce performance.
• Working storage: Short‑term use at 4°C for hours to a few days is acceptable; for >1–2 weeks, store at −20°C; for long‑term, follow product specification of −80°C.
• Buffer: If reconstitution is required (lyophilized material), dissolve in nuclease‑free water or TE (pH 7.5–8.0). Final buffer composition and recommended concentration are not specified for this item; refer to CoA/Spec Sheet.
• Avoid: Multiple freeze–thaw cycles, nucleases, and low‑pH conditions. Use sterile, low‑binding tips and tubes.
• Documentation: Record lot number, aliquot map, and calculated concentrations to ensure traceability for downstream experiments.

Always consult the CoA/Spec Sheet for any lot‑specific instructions.

Structure and Identity

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

What this product is (general/literature description):

• pLenti-YTHDC2-sgRNA denotes a lentiviral vector plasmid carrying a single-guide RNA (sgRNA) designed to target the YTHDC2 gene. As named, it is a circular double‑stranded DNA construct intended for CRISPR workflows and lentiviral packaging.
• Typical components of lentiviral sgRNA plasmids (literature/general): U6 or similar RNA polymerase III promoter driving the sgRNA, a guide scaffold, and lentiviral long terminal repeats (LTRs)/packaging signals enabling production of replication‑incompetent lentiviral particles when co-transfected with helper plasmids.
• Structural features (general):
  • Macromolecule class: nucleic acid (plasmid DNA, circular dsDNA).
  • Functional elements may include antibiotic resistance cassette(s), an origin of replication, and selectable markers; specific features for this SKU are not specified.

Molecular formula/MW: Not applicable to heterogeneous plasmid DNA and not specified for this item.

2D/architectural description (general): The vector backbone forms a closed circular dsDNA; functional modules (promoter, guide scaffold, selection marker, LTRs, Psi packaging signal) are arranged in a standard lentiviral topology, permitting expression of an sgRNA and packaging into viral particles when supplied with trans‑acting gag/pol and env from helper constructs.

Synthetic Utility

Chemical synthesis utility is not applicable; however, the construct provides substantial “synthetic biology” utility.

General/literature perspective on plasmid utility:

• Modular cloning: Lentiviral sgRNA vectors are amenable to Golden Gate, Gibson, or classical ligation to insert target‑specific protospacers into the sgRNA expression cassette.
• Vector engineering: Backbones typically allow swapping of promoters, selection markers, and addition of barcodes for pooled screens. Such modifications enable tailored expression and tracking in complex experiments.
• Combinatorial perturbations: Co‑expression with Cas9 or dCas9 fusions (KRAB, VP64, p300) facilitates gene knockdown/activation screens; dual‑gRNA architectures enable deletions or multiplexed editing.
• Retrosynthetic planning (bio): Starting from a validated pLenti sgRNA backbone, one can introduce alternative protospacers targeting different exons of YTHDC2 or orthologs, or adapt PAM requirements for distinct Cas variants (SpCas9, SaCas9, Cas12a), depending on experimental needs.

Item-specific cloning sites, restriction map, and antibiotic resistance markers are not provided; refer to CoA/Spec Sheet for exact backbone features.

Target Specificity

Only information directly from the Product Data may be provided here.

• Target indicated by product name: YTHDC2 (gene).
• Guide sequence, PAM, exon/intron location, and species reactivity: Not specified for this item; refer to CoA/Spec Sheet.
• Clone/backbone identity, selection marker, and reporter elements: Not specified for this item; refer to CoA/Spec Sheet.

Note: Despite the Category Path labeling under antibodies, this SKU denotes a plasmid sgRNA reagent by name; treat as a genetic tool rather than an antibody.

Preguntas frecuentes

How should this product be stored?

Store at ?80 °C. Ultra-low temperature storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships frozen with dry ice and cold packs. Unpack on arrival and transfer it to the storage condition stated above; handle dry ice with insulated gloves in a ventilated area.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.

Which species reactivity has this antibody been validated against?

Validated reactivity is listed on the PDP and on each lot's datasheet. We validate against the most commonly tested species for the target (pLenti-YTHDC2-sgRNA); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.