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This reference provides a concise overview of commonly used in vivo AAV delivery routes in mice, including typical injection volumes, vector doses, target tissues, and expected expression timelines. The values presented represent standard experimental ranges and may vary depending on factors such as AAV serotype, promoter selection, vector quality, animal age and strain, and specific study objectives.
🧠 CNS — Direct Brain & CSF Delivery
Intracerebral (Stereotaxic)
Use: Precise targeting of defined brain regions
- Volume: 100 ~ 500 nL per site (typical 100 ~ 300 nL)
- Dose: 10⁹ ~ 10¹¹ VG/site (up to ~10¹²)
- Onset: 3 ~ 7 days
- Robust: 3 ~ 4 weeks
- Duration: Months–lifetime (neurons)
- Highest spatial precision
- Minimal peripheral exposure
Intracerebroventricular (ICV)
Use: Broad CNS distribution via CSF
- Volume: Adult 2 ~ 5 µL • Neonatal 1 ~ 2 µL
- Dose: 10¹⁰ ~ 10¹² VG total
- Onset: 4 ~ 7 days
- Robust: 3 ~ 4 weeks
- Neonatal ICV: near brain-wide transduction
Intrathecal (IT) — Lumbar or Cisterna Magna
Use: Spinal cord ± brain via CSF
- Volume: 5 ~ 10 µL
- Dose: 10¹¹ ~ 10¹² VG
- Onset: ~1 week
- Robust: 3 ~ 4 weeks
- Less invasive than intracranial surgery
❤️ Systemic Delivery (Intravenous)
Tail Vein IV / Retro-Orbital IV / Neonatal Facial Vein
Use: Whole-body delivery (tropism = capsid-dependent)
Routes included:
- Tail vein (adult)
- Retro-orbital sinus (adult)
- Facial vein (neonate)
Volume:
- Tail vein: 100 ~ 200 µL
- Retro-orbital: 100 ~ 150 µL (up to ~200 µL adult)
- Facial vein (neonate): 20 ~ 50 µL
- Dose: 10¹¹ ~ 10¹³ VG per mouse
Expression:
- Peripheral tissues: 5 ~ 10 days
- CNS with BBB-crossing capsids: 3 ~ 4 weeks
- Peak: ~4 weeks
- Retro-orbital often easiest and most reliable IV method
- Tail vein requires more technical skill
Important: Retro-orbital injection is IV access via the venous sinus — it does not target the eye.
💪 Peripheral Local Delivery
Intramuscular (IM)
Use: Muscle expression or systemic secretion
- Volume: 10 ~ 50 µL per muscle
- Dose: 10¹⁰ ~ 10¹² VG/site
- Onset: ~1 week
- Robust: 3 ~ 4 weeks
- Duration: Very long-term
- Muscle is highly permissive for AAV
Intraocular — Intravitreal / Subretinal
Use: Retina and visual system
Intravitreal
- Volume: 1 ~ 2 µL
- Targets: Ganglion cells
Subretinal
- Volume: 0.5 ~ 1 µL
- Targets: Photoreceptors/RPE
Dose: ~10⁹ ~ 10¹⁰ VG/eye
Expression: Onset 1 ~ 2 weeks • Peak 3 ~ 4 weeks
🟡 Adipose Tissue — Brown, White & Visceral Fat Delivery
AAV-mediated gene delivery to adipose tissue can be performed by direct injection into individual fat depots, intraperitoneal (IP) administration, or systemic intravenous (IV) delivery. The optimal dose depends strongly on the AAV capsid, targeted adipose depot, administration route, and desired degree of tissue specificity.
Engineered adipose-tropic capsids such as AAV-Rec2 and related variants generally require substantially lower doses than conventional systemic AAV serotypes.
Intraperitoneal (IP) — Visceral Adipose Tissue
Use: Broad targeting of visceral white adipose tissue (VAT), including epididymal/gonadal, mesenteric, and retroperitoneal fat
- Recommended Dose: ~1 × 10¹⁰ – 4 × 10¹⁰ VG/mouse
- Typical Volume: 100–200 µL
- Preferred Capsids: Adipose-tropic capsids such as Rec2 or related engineered variants
- Targets: eWAT/gWAT, mesenteric WAT, retroperitoneal WAT and other visceral fat depots
- Expression: Detectable within ~1–2 weeks; robust expression typically by ~2–4 weeks
- Particularly useful when multiple visceral fat depots need to be targeted simultaneously
- Off-target liver transduction depends strongly on capsid and vector design
Published studies using Rec2 have demonstrated efficient visceral adipose transduction at approximately 1–2 × 10¹⁰ VG/mouse following IP administration.
Direct Adipose Depot Injection
Use: Localized gene delivery to a specific brown or white adipose tissue depot
Brown Adipose Tissue (BAT)
- Typical Dose: ~1 × 10⁹ – 2 × 10¹⁰ VG/depot
- Target: Interscapular BAT
- Allows highly localized manipulation of brown adipose tissue while minimizing systemic exposure
White Adipose Tissue (WAT)
- Typical Dose: ~1 × 10¹⁰ – 2 × 10¹¹ VG/depot
- Targets: Inguinal WAT (iWAT), epididymal/gonadal WAT (eWAT/gWAT), or other accessible fat pads
- Higher doses may be required depending on capsid, depot size, and desired transduction efficiency
For highly adipose-tropic capsids such as Rec2, effective direct-depot transduction has been reported at approximately 1 × 10⁹ VG for BAT and 1 × 10¹⁰ VG for iWAT.
Systemic IV — Adipose Tissue Targeting
Use: Broad adipose targeting when local or IP administration is not appropriate
- Typical Dose: ~1 × 10¹² – 5 × 10¹² VG/mouse
- Routes: Tail vein or retro-orbital IV
- Common Serotypes: AAV8, AAV9, or engineered adipose-tropic capsids
- Conventional AAV8/AAV9 generally require substantially higher systemic doses than adipose-tropic engineered capsids
- Liver and other peripheral tissues may receive substantial vector exposure
- Tissue-specific promoters and/or miRNA-based detargeting strategies can improve adipose specificity
Practical AAV Dose Guide for Mouse Adipose Studies
| Administration Route | Target | Typical AAV Dose |
|---|---|---|
| IP — adipose-tropic capsid | Visceral WAT | 1 × 10¹⁰ – 4 × 10¹⁰ VG/mouse |
| Direct BAT injection | Brown adipose tissue | 1 × 10⁹ – 2 × 10¹⁰ VG/depot |
| Direct WAT injection | iWAT/eWAT/gWAT | 1 × 10¹⁰ – 2 × 10¹¹ VG/depot |
| Systemic IV | Multiple adipose depots | 1 × 10¹² – 5 × 10¹² VG/mouse |
Important: These values should be considered starting ranges rather than fixed doses. Optimal dosing depends on the AAV capsid, promoter, transgene, mouse strain and age, adipose depot, vector purity, and experimental objective. Adipose-tropic engineered capsids may achieve efficient transduction at substantially lower doses than conventional AAV8 or AAV9.
⚙️ Additional Common Routes
Intraperitoneal (IP)
Use: Neonatal systemic delivery or peritoneal targeting
- Volume: 50 ~ 200 µL
- Dose: 10¹¹ ~ 10¹³ VG
- Less efficient than IV
Intranasal
Use: Respiratory tract ± olfactory CNS entry
- Volume: 10 ~ 30 µL (dropwise)
- Dose: 10¹⁰ ~ 10¹¹ VG
- Variable brain delivery
Intrahepatic (Neonatal)
Use: Liver-directed gene transfer
- Volume: 20 ~ 50 µL
- Highly efficient hepatic transduction
🧾 Quick Route Selection Guide
| Route | Target Tissues | Typical Volume | Typical Dose (VG/mouse) | Onset of Expression | Peak Expression | Duration |
|---|---|---|---|---|---|---|
| Tail Vein (IV) | Liver (strong), heart, muscle, systemic | 100 ~ 200 µL | 1×10¹¹ ~ 5×10¹² | 3 ~ 7 days | 3 ~ 4 weeks | Months to lifetime |
| Retro-Orbital (IV) | Systemic (similar to tail vein) | 100 ~ 150 µL | 1×10¹¹ ~ 5×10¹² | 3 ~ 7 days | 3 ~ 4 weeks | Months to lifetime |
| Neonatal Facial Vein (IV) | Widespread systemic (neonates) | 20 ~ 50 µL | 1×10¹⁰ ~ 5×10¹¹ | 3~ 5 days | 3 ~ 4weeks | Long-term |
| Intracerebral (Stereotaxic) | Local brain region | 100 ~ 500 nL | 1×10⁹ ~ 1×10¹¹ | 1 ~ 2 weeks | 3 ~ 5 weeks | Long-term |
| Intracerebroventricular (ICV) | CNS (widespread, esp. neonates) | Adult: 5 ~ 10 µL Neonate: 2 ~ 4 µL | 1×10¹⁰ ~ 1×10¹² | 1~ 2 weeks | 3 ~ 6 weeks | Long-term |
| Intrathecal (IT) (lumbar or cisterna magna) | Spinal cord, DRG, CNS | 5 ~ 10 µL | 1×10¹⁰ ~ 1×10¹² | 1 ~ 2 weeks | 3 ~ 5 weeks | Long-term |
| Intramuscular (IM) | Injected muscle, motor neurons (retrograde) | 10 ~ 50 µL | 1×10¹⁰ ~ 1×10¹² | 1 ~ 2 weeks | 3 ~ 4 weeks | Months to lifetime |
| Intraperitoneal (IP) | Peritoneal organs, liver (moderate systemic) | 200 ~ 500 µL | 1×10¹¹ ~ 5×10¹² | 1 ~ 2 weeks | 3~ 5 weeks | Long-term |
| Subcutaneous (SC) | Local tissue, lymphatics | 50 ~ 200 µL | 1×10¹⁰ ~ 1×10¹² | 1 ~ 2 weeks | 3 ~ 5 weeks | Long-term |
⏱️ General AAV Expression Kinetics (Across Routes)
Typical single-stranded AAV:
- Early expression: 3 ~ 7 days
- Robust expression: 3 ~ 4 weeks
- Peak: ~4 weeks
- Duration: Months–years in non-dividing tissues
Faster onset with:
- Highly permissive tissues (muscle, liver)
- Self-complementary AAV (scAAV)
- Strong promoters (CMV, CAG)
- Higher MOI
🧪 Key Factors That Shift These Numbers
- Serotype (e.g., AAV9 vs AAV8 vs PHP.eB)
- Promoter strength (CMV vs CAG vs tissue-specific)
- Vector genome form (ssAAV vs scAAV — self-complementary expresses faster)
- Age of animal (neonatal transduction is typically broader and faster)
- Immune status
- Purity / empty capsids
- Injection accuracy
⭐ Practical Notes for Experimental Design
- Pain / sensory research: Intrathecal injection
- Systemic liver targeting: Tail vein or retro-orbital IV with AAV8/AAV9
- Whole-brain delivery: Neonatal ICV or systemic PHP.eB (mouse strain-dependent)
- Region-specific neuroscience studies: Stereotaxic intracerebral injection
- Motor neuron studies: Intramuscular with retrograde serotypes
Still unsure about the optimal AAV administration route? Please consult our Ph.D scientists for expert guidance
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