AAV vectors can be engineered to support multiple RNA-based strategies for precise regulation of gene expression. In addition to transgene overexpression, our AAV vector design platform enables expression of shRNA for RNAi-mediated gene silencing, shRNAmir constructs for regulated or tissue-specific knockdown, and miRNA systems for studying endogenous post-transcriptional regulation. The platform also supports gRNA expression for CRISPR-based genome editing or transcriptional modulation, as well as Tough Decoy (TuD) RNA for effective inhibition of specific microRNAs. Through optimized vector architecture and promoter selection (Pol II or Pol III), these systems enable efficient and stable RNA expression for gene regulation and genome engineering applications in vitro and in vivo.

shRNA: Enables robust and stable gene knockdown through RNA interference (RNAi) by producing a hairpin RNA that is processed into siRNA, leading to targeted degradation of the corresponding mRNA. Leveraging a proprietary database of over 80,000 pre-validated target regions covering both human and mouse genes, we can rapidly identify high-confidence shRNA sequences with strong and reproducible knockdown performance.

shRNAmir: Allows regulated or tissue-specific gene silencing by embedding the shRNA sequence within a microRNA scaffold, enabling expression from Pol II promoters and processing through the endogenous miRNA biogenesis pathway for efficient and physiologically compatible gene knockdown.

miRNA: Facilitates modulation of endogenous gene expression networks by expressing microRNAs that regulate target mRNAs through translational repression or mRNA destabilization. Precursor miRNA sequences are cloned to ensure proper processing and efficient expression of mature miRNAs.

gRNA: Enables sequence-specific targeting for CRISPR systems by guiding Cas nucleases or dCas-based transcriptional regulators to defined genomic loci through complementary base pairing. This allows precise genome editing, gene activation, gene repression, or epigenetic modulation depending on the CRISPR effector used, making gRNA a central component for programmable genome engineering and functional genomics studies.

TuD RNA: Enables potent and durable inhibition of specific microRNAs through engineered tough decoy structures that bind and sequester mature miRNAs, preventing them from interacting with their endogenous targets. To maximize inhibition efficiency, dual miRNA binding sites (2× MBS) are designed and arranged in an antiparallel orientation.