Development of an adipose-tropic AAV capsid ablating liver tropism

AAV vectors are mainstream delivery platforms in gene therapy, yet AAV-mediated gene transfer to adipose tissue is underdeveloped due to low efficiency of natural AAVs. We previously demonstrated that an engineered capsid Rec2 displayed improved adipo-tropism but with the caveat of liver transduction. To generate highly adipo-tropic capsid, we modified Rec2 capsid by site-specific mutagenesis and found the variant V7 with F503Y, Y708D and K709I substitution to harbor highly selective adipo-tropism while diminishing liver transduction. Intraperitoneal injection favored transduction to visceral fat while intravenous administration favored subcutaneous fat.  Intraperitoneal administration of V7 vector harboring human leptin and adiponectin as single transcript normalized the metabolic dysfunction of ob/ob mice at a low dose. Moreover, introducing the same mutagenesis to AAV8 capsid diminished liver transduction suggesting F503, Y708 and K709 critical for liver transduction. The Rec2.V..., Rec2 capsid V7 mutagenesis
The Rec2 capsid was mutagenized at sites indicated in Figure1A and full capsid sequence is listed in Figure 1B. We name Rec2 variants as V1 to V8 in numerical order. GeneArt site-directed mutagenesis Plus kit (A14604, Invitrogen) was used to carry out the designated mutagenesis.  Rec2 capsid sequence is based on published data6. Primers for mutagenesis was synthesized by Sigma-Aldrich: F503Y (ACAACAATAGCAACTATGCCTGGACTGCTGG,CCAGCAGTCCAGGCATAGTTGCTATTGTTGT); Y708D and K709I (TACACCTCCAACTACGACATATCTACAAGTGTGGA, TCCACACTTGTAGATATGTCGTAGTTGGAGGTGTA).
Mutagenesis was set up and carried out according to the manufacturer’s instruction, and the nucleotide replacement at each site was confirmed by sequencing at the Core Facility of The Ohio State University, Comprehensive Cancer Center. The whole capsid was also sequenced and confirmed without unintended mutation.
AAV8 capsid mutation
Rec2 capsid was generated from capsid domain shuffling between rh20 and AAV8 6 ..., , # Data from: Development of an adipose-tropic AAV capsid ablating liver tropism

[https://doi.org/10.5061/dryad.cjsxksnh6](https://doi.org/10.5061/dryad.cjsxksnh6)

## Description of the data and file structure

Raw data for the paper entitled \"Development of an adipose-tropic AAV capsid ablating liver tropism\"

AAV vectors targeting adipose tissue are still underdeveloped. The F503Y/Y708D/K709I substitution in AAV8 and Rec2 capsids eliminates liver tropism, while the Rec2 variant V7 with this substitution is highly adipo-tropic. V7 vector-based gene therapy effectively rescues metabolic defects in ob/ob mice at a low dose. Tissue types: epididymal (male) or gonadal (female) white adipose tissue (eWAT or gWAT), mesentery white adipose tissue (mWAT), retroperitoneal white adipose tissue (rWAT), inguinal white adipose tissue (iWAT) and intrascapular brown adipose tissue (BAT).

### Files and variables

#### File: V7\_Fig2\_Data.csv

**Description:** raw data for Fig 2

##### Variables

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