AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis

Targeted gene therapy has shown great promise for treating rare diseases, particularly those that arise from single gene defects. Recombinant adeno-associated virus (AAV) has recently emerged as a highly popular gene delivery vector for a wide range of gene therapy and vaccine applications. One essential consideration when evaluating the therapeutic potential and safety profile of an AAV vector is its distribution and persistence in various organs and tissues. While qPCR-based methods provide average values for copy number, the RNA in situ hybridization (ISH) technology provides morphology-based, cell-specific quantification of vector DNA, transgene mRNA, and specific biomarkers.

As stated by the USFDA, "The biodistribution data, coupled with other preclinical safety endpoints such as clinical pathology and histopathology, help determine whether vector presence or gene expression correlates with any tissue-specific detrimental effects in animals." (FDA Guidance for Industry: Preclinical Assessment of Investigational Cellular and Gene Therapy Products, November 2013). This is particularly important in germ-line tissues (i.e., gonads) to determine intertied transmission status. Our AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis service can help you validate and accelerate your gene therapy development.

The RNA ISH technology is an excellent solution for detecting AAV vector DNA and therapeutic transgene mRNA expression within intact tissue morphology, addressing critical questions on tissue biodistribution, persistence, cellular tropism, and vector promoter activity. As a leading technology provider, Creative Bioarray's elite scientists have broad experience in AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis (RNA ISH method), which can supply the best service for you.

Figure 1. Visualization of AAV-transduced cells and specific cell types in the monkey retina. (A) The AAV vector used in this study contains a GFP transgene under the control of the CB promoter. RNAscope® probes were designed to target the CB promoter DNA sequence (V-CB promoter) as well as the GFP transgene mRNA (GFP-C2). (B) RNAscope® probes were used to detect the rod cells marked by Rhodopsin (Mfa-RHO) and cone cells marked by Opsin 1 short wave sensitive (Mfa-OPN1SW-C2).

Features of AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis (RNA ISH method)

(1) Detection of single-copy DNA vectors
(2) Codon-optimized transgene and biomarker expression analysis
(3) Custom probes designed within 1-2 weeks
(4) Fastest turnaround time

Benefits of AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis (RNA ISH method)

(1) Simultaneously visualize in vivo delivery of vector DNA and therapeutic transgene mRNA
(2) Identify cell tropism of AAV vector by multiplexing with cell-type markers
(3) Distinguish transgene from endogenous sequences, at the single nucleotide level
(4) Quantify AAV+ cell number and track persistence over time

Creative Bioarray offers AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis for you as follows:

• Probe design
• Probe synthesis
• ISH staining
• Imaging
• Data analysis

Quotation and ordering

Our customer service representatives are available 24hr a day! We thank you for considering Creative Bioarray as your AAV Vector/Transgene/Biomarker RNA Biodistribution Analysis partner.

References

1. Hastie E and Samulski RJ. Adeno-associated virus at 50: A golden anniversary of discovery, research, and gene therapy success — a personal perspective. Hum Gene Ther. 2015; 26:257–265
2. Wang F, et al. RNAscope®: A novel in situ RNA analysis platform for formalin-fixed, paraffinembedded tissues. J Mol Diagn. 2012; 14(1):22–9.
3. Polinski NK, et al. Impact of age and vector construct on striatal and nigral transgene expression. Mol Ther Methods Clin Dev. 2016; 3, 16082
4. Keeler AM, et al. Cellular analysis of silencing the Huntington's Disease gene using AAV9 mediated delivery of artificial microRNA into the striatum of Q140/Q140 mice. J Huntingtons Dis. 2016; 5(3):239–248
5. Borel F, et al. Therapeutic rAAVrh10 mediated SOD1 silencing in adult SOD1G93A mice and nonhuman primates. Hum Gene Ther. 2016; 27(1):19–31.