pLenti-YTHDC2-sgRNA

Cat. No.: P751892

Specifications

Product Name

pLenti-YTHDC2-sgRNA

Product Line

Biological Products

Storage and Shipping

Storage

Store at -80°C

Shipped In

Dry ice packs + Cold packs

Stability And Storage

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

Application Protocols

Item-specific tested applications, conditions, and positive-control details are Not specified for this item; refer to CoA/Spec Sheet.

General literature guidance (not product-validated):

• For transient plasmid delivery, follow the transfection reagent’s protocol for DNA amount per well/plate format and cell type.
• For lentiviral workflows, follow BSL-2 SOPs for packaging, titering, and transduction. Validate editing via sequencing-based assays and include appropriate controls.

Because application conditions vary widely by cell line and delivery method, users should optimize empirically.

Biological Roles

YTHDC2 (YTH domain-containing protein 2) is an RNA helicase with a YTH domain that recognizes N6-methyladenosine (m6A)-modified RNA. The following information is general literature context for the target gene; it is not specific to this product’s guide design or efficacy:

• Molecular function (literature): RNA binding via YTH domain; ATP-dependent RNA helicase activity; implicated in regulation of mRNA stability and translation of m6A-marked transcripts.
• Biological processes (literature): Plays roles in germ cell development, meiotic progression, and broader post-transcriptional regulation pathways.
• Cellular localization (literature): Predominantly cytoplasmic with association to RNA-protein granules, depending on cell type and context.
• Pathway context: Interfaces with the m6A epitranscriptomic machinery together with readers (YTH family), writers (METTL3/METTL14 complex), and erasers (FTO/ALKBH5).

Implication for research: Perturbation of YTHDC2 via CRISPR knockout/knockdown can be used to study RNA metabolism, m6A-dependent translation control, and meiosis-related gene regulation. Species specificity of this product’s sgRNA, target exon/intron, and off-target risk are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

This item is a plasmid DNA reagent and is not used as a chemical buffering agent.

• Working solutions (general, literature): For storage and handling, nuclease-free water or TE buffer (10 mM Tris·Cl, 0.1–1 mM EDTA, pH 7.5–8.5) are commonly used to stabilize plasmid DNA. Avoid divalent cations that can promote nuclease activity.

No electrophoresis running buffers or cell culture media formulations are specific to this product; use standard molecular biology buffers appropriate to your downstream workflow.

Green Alternatives

Classical “green solvent” substitutions are not applicable to plasmid DNA reagents. Nevertheless, greener and safer laboratory practices can be considered (general guidance):

• Reduce hazardous solvent use: Prefer column-free plasmid cleanup kits or aqueous-based endotoxin removal where feasible; minimize phenol/chloroform extractions.
• Minimize single-use plastics: Use reusable racks and recycle eligible plastics in compliance with biosafety rules.
• Safer delivery options: For transient studies not requiring genomic integration, consider non-viral delivery (e.g., electroporation or lipid-mediated transfection) to reduce the need for viral packaging and associated BSL-2 procedures. This is a workflow alternative, not a substitute product.
• Energy efficiency: Consolidate −80°C storage and use high-efficiency freezers to reduce energy footprint.

These are general sustainability practices and not item-specific specifications.

Pharmaceutical Uses

Not applicable. This product is for research use only and is not intended for human or veterinary use, diagnostics, or as a pharmaceutical ingredient.

General context (literature): Plasmid and viral vector technologies may be used in bioproduction research and process development settings for gene modulation studies. However, any clinical, diagnostic, or GMP manufacturing applications require purpose-built materials and regulatory oversight. This item provides no pharmacopeial status or excipient role.

Item-specific regulatory/quality designations: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Nucleic-acid constructs do not have fixed physicochemical constants like small molecules. Only general/literature characteristics are provided; they are not item-specific specifications.

• Physical state: Aqueous DNA solution or lyophilized DNA (item-specific appearance not specified).
• Molecular weight/length: Not specified for this item; refer to CoA/Spec Sheet. (Typical plasmids: ~6–12 kb; literature)
• Solubility: Highly soluble in nuclease-free water and low-salt buffers (e.g., 10 mM Tris·Cl, pH 7.5–8.5; literature).
• pH stability (literature): Stable near neutral to slightly alkaline pH; prolonged exposure to acidic pH promotes depurination.
• Temperature sensitivity: DNA integrity is preserved at ≤−20°C; optimal long-term storage at −80°C (item requires −80°C; see Storage & Reconstitution).
• UV characteristics (literature): A260 for quantification; A260/A280 ~1.8 indicates low protein contamination; A260/A230 >2.0 indicates low salt/organic carryover.
• DNase/RNase sensitivity: Susceptible to nuclease degradation; maintain nuclease-free conditions (literature).

No boiling/melting point, density, refractive index, logP, or pKa are applicable to plasmid DNA. For any lot-specific concentration, buffer composition, or OD ratios: Not specified for this item; refer to CoA/Spec Sheet.

Quality & Grades

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Typical quality attributes for plasmid/CRISPR reagents (general, literature):
  • Sequence verification of the sgRNA spacer and backbone junctions.
  • Plasmid integrity confirmed by restriction digest and/or full plasmid sequencing.
  • Endotoxin level control for transfection into sensitive cell types (e.g., <0.1 EU/µg for some applications) — item-specific value not specified.
  • Sterility and absence of mycoplasma (where applicable) — not specified for this item.
• Implications:
  • High-purity, sequence-verified plasmids improve cloning success, transfection efficiency, and the fidelity of CRISPR targeting.
  • Endotoxin-reduced preparations are preferred for primary cells and in vivo model research (research use only).
• Documentation: For lot-specific QC metrics (concentration, A260/280, sequence confirmation, endotoxin), Not specified for this item; refer to CoA/Spec Sheet.

Note: This item’s category path lists “抗体 (antibody),” but the product name indicates a pLenti sgRNA plasmid. Users should rely on the item’s CoA/Spec Sheet for definitive format and QC details.

Reaction & Applications

As a pLenti-based CRISPR sgRNA construct targeting YTHDC2, this reagent supports gene perturbation workflows. The following are general research applications (literature), not item-specific claims:

• CRISPR-Cas9 knockout: Express the sgRNA with a nuclease-active Cas9 (from a co-transfected plasmid, stable line, or lentiviral co-delivery) to induce indels at the YTHDC2 locus via non-homologous end joining.
• CRISPRi/CRISPRa: When paired with dCas9-KRAB or dCas9-VP64/SunTag systems, the same sgRNA architecture (with promoter-appropriate expression) can mediate transcriptional repression or activation if guides are positioned near TSS (guide position not provided here).
• Stable cell line generation: Use lentiviral packaging (e.g., 2nd/3rd generation systems) to integrate the sgRNA cassette for long-term studies; select with the backbone’s resistance marker (if present; not specified for this item).
• Functional genomics: Targeted loss-of-function screens, epistasis mapping, pathway analysis, and RNA metabolism studies related to YTHDC2 (an m6A-associated helicase; see Biological Roles).
• Validation: Employ T7E1, Surveyor, Inference of CRISPR Edits (ICE), or NGS amplicon sequencing to quantify editing efficiency (general best practices).

Note: Guide sequence, PAM context, species/isoform targeting, and selectable markers are Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

While not a chemical reaction reagent, typical operational conditions for workflows using pLenti sgRNA constructs are summarized here as general literature guidance (not item-specific):

• Transfection (plasmid): HEK293T or similar cells, 70–90% confluency; DNA:transfection reagent ratios per vendor instructions; incubate 24–72 h before analysis.
• Lentiviral packaging: Co-transfect pLenti-sgRNA with packaging plasmids (e.g., psPAX2, pMD2.G or system equivalents). Harvest viral supernatant at 48–72 h; filter (0.45 µm). Perform under BSL-2.
• Transduction: Add viral supernatant with polybrene (if compatible) to target cells; spinoculation may enhance efficiency. Select with antibiotic marker encoded by the vector (marker for this item not specified).
• Editing assessment: Allow 48–96 h post-delivery before measuring indels; for stable pools/isolates, expand 1–2 weeks with selection.
• Controls: Include non-targeting sgRNA and positive-control sgRNA; verify on-target activity by amplicon sequencing or ICE/TIDE.

Exact MOI, selection agent and dose, guide sequence, PAM context, and promoter details are Not specified for this item; refer to CoA/Spec Sheet and institutional SOPs.

Safety & Handling

• Research Use: For research use only.
• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Biosafety (general, literature):
  • pLenti backbones are used to generate replication-incompetent lentiviral particles; activities involving packaging and transduction are typically conducted under BSL-2 containment with appropriate approvals and training.
  • Use biological safety cabinets (Class II) for handling lentiviral packaging/transduction mixtures; decontaminate with fresh 0.5–1% sodium hypochlorite or appropriate disinfectant.
• Handling practices:
  • Work in nuclease-free conditions (RNase-/DNase-free tubes, tips, and buffers) to maintain integrity of the sgRNA cassette.
  • Avoid repeated freeze–thaw; keep on ice during use.
• Incompatibilities (general): Strong acids/alkalis and nucleases degrade DNA; avoid microbial contamination.
• PPE: Lab coat, gloves, eye protection. When aerosol-generating steps or viral packaging are involved, use additional BSL-2 PPE per institutional policy.
• First aid (general): In case of exposure during viral work, follow institutional exposure protocols; wash affected area with soap/water, flush eyes with water, and seek medical evaluation per EHS guidance.

Always consult the product SDS and institutional biosafety office for authoritative safety controls and waste disposal.

Solvent Selection

This is a DNA plasmid reagent; conventional solvent selection frameworks for small-molecule solutes do not apply.

• Practical medium considerations (general, literature):
  • Compatible diluents: Nuclease-free water or low-ionic-strength buffers (e.g., 10 mM Tris·Cl, pH 7.5–8.5; 0.1–1 mM EDTA may be included to chelate divalent cations and reduce nuclease activity).
  • Avoid: Phenol, chloroform, and alcohol residues in working stocks; high salt may inhibit downstream transfection/enzymatic reactions.
  • For transfection: Follow the transfection reagent/vendor recommendations regarding buffer composition and DNA concentration.

If this item is supplied lyophilized or as a concentrated stock, reconstitute/dilute in nuclease-free water or TE buffer unless otherwise specified in the CoA/Spec Sheet (item-specific instructions not provided here).

Storage & Reconstitution

• Storage conditions (item-specific): Store at −80°C.
• Shipping (item-specific): Shipped on dry ice packs + cold packs.
• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Reconstitution (general guidance, if lyophilized or concentrated):
  • Use nuclease-free water or TE buffer (10 mM Tris·Cl, 0.1–1 mM EDTA, pH 7.5–8.5).
  • Mix gently by pipetting; avoid vigorous vortexing that can shear DNA.
  • Determine concentration by A260 or fluorometric assay; store working aliquots to minimize freeze–thaw.
• Handling:
  • Keep on ice during use; return promptly to −80°C.
  • Use low-bind, nuclease-free tubes and filter tips.
• Stability: Long-term stability is maximized at −80°C. Avoid repeated freeze–thaw cycles. Lot-specific stability or expiration: Not specified for this item; refer to CoA/Spec Sheet.

Follow any additional instructions provided with the specific lot documentation.

Structure & Identity

This product is a lentiviral CRISPR plasmid construct expressing a single guide RNA (sgRNA) designed to target the YTHDC2 gene (pLenti-YTHDC2-sgRNA). It is a nucleic-acid-based research tool rather than a small-molecule chemical.

• Item-specific identifiers:
  • CAS: Not applicable
  • SMILES / InChI / InChIKey: Not applicable to plasmid DNA
  • Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
• Structural features (general, literature):
  • Backbone: pLenti (lentiviral) plasmid backbone enabling packaging into lentiviral particles for stable integration in dividing and non-dividing cells.
  • Expression cassettes: Typically include a U6 or H1 promoter for sgRNA expression and may include antibiotic selection and/or reporter markers (literature; item-specific features not specified here).
  • Guide architecture: Standard CRISPR sgRNA scaffold downstream of a polymerase III promoter (literature).
• 2D/functional description (general): circular double-stranded DNA plasmid; functional modules comprise origin(s) of replication, selection markers, promoter–sgRNA cassette, and lentiviral long terminal repeats (LTRs) for packaging (literature).

Synthetic Utility

In the context of molecular biology (rather than small-molecule synthesis), pLenti-YTHDC2-sgRNA provides utility for constructing gene perturbation systems and genetic circuits.

• Modular cloning (general, literature): The sgRNA spacer can be exchanged by Golden Gate or annealed oligo cloning into a BsmBI/BbsI site in many CRISPR backbones (vector-specific; sites for this item not specified).
• Genome engineering workflows:
  • Knockout via Cas9: Introduce frameshifts at YTHDC2 coding regions.
  • CRISPRi/a: Repress or activate YTHDC2 transcription with dCas9 fusions when guides are placed near the TSS.
• Combinatorial studies: Co-express multiple sgRNAs for multiplex perturbation of YTHDC2 with interacting factors (requires suitable backbone/expressions modules; not specified for this item).
• Readouts: Use qPCR, western blot, RNA-seq, RIP-seq, or CLIP-based assays to assess YTHDC2 expression changes and downstream RNA metabolism (general guidance).

Backbone features such as antibiotic resistance, origin of replication, and promoter elements are Not specified for this item; refer to CoA/Spec Sheet for cloning and propagation details.

Target Specificity

• Intended target: YTHDC2 gene (guide sequence not provided).
• Species/isoform: Not specified for this item; refer to CoA/Spec Sheet.
• Off-target information: Not specified for this item; refer to CoA/Spec Sheet.
• Validation data (editing efficiency, on-target cut site): Not specified for this item; refer to CoA/Spec Sheet.

Only product-data-derived details can be listed here; since none are provided, users should consult the CoA/Spec Sheet for guide spacer sequence, PAM, strand orientation, genomic coordinates, and any tested cell lines.