Version 1
: Received: 2 August 2024 / Approved: 2 August 2024 / Online: 5 August 2024 (07:11:58 CEST)
How to cite:
Boychenko, S.; Abdullina, A.; Laktyushkin, V.; Brovin, A.; Egorov, A. Assessment of Adipocyte Transduction Using Different AAV Capsid Variants. Preprints2024, 2024080199. https://doi.org/10.20944/preprints202408.0199.v1
Boychenko, S.; Abdullina, A.; Laktyushkin, V.; Brovin, A.; Egorov, A. Assessment of Adipocyte Transduction Using Different AAV Capsid Variants. Preprints 2024, 2024080199. https://doi.org/10.20944/preprints202408.0199.v1
Boychenko, S.; Abdullina, A.; Laktyushkin, V.; Brovin, A.; Egorov, A. Assessment of Adipocyte Transduction Using Different AAV Capsid Variants. Preprints2024, 2024080199. https://doi.org/10.20944/preprints202408.0199.v1
APA Style
Boychenko, S., Abdullina, A., Laktyushkin, V., Brovin, A., & Egorov, A. (2024). Assessment of Adipocyte Transduction Using Different AAV Capsid Variants. Preprints. https://doi.org/10.20944/preprints202408.0199.v1
Chicago/Turabian Style
Boychenko, S., Andrew Brovin and Alexander Egorov. 2024 "Assessment of Adipocyte Transduction Using Different AAV Capsid Variants" Preprints. https://doi.org/10.20944/preprints202408.0199.v1
Abstract
Adeno-associated viruses (AAV) are widely used as viral vectors for gene delivery in mammalian cells. We focused on the efficacy of transduction of AAV 2/5, 2/6, 2/8 and 2/9 expressing GFP in 3T3-L1 murine preadipocyte cells by live imaging microscopy using IncuCyte S3 and flow cytom-etry. Three transduction modes were assessed: AAV transduction in 3T3-L1 preadipocyte cells with or without further differentiation into mature adipocyte-like cells and injection of AAVs in differ-entiated adipocyte-like cells. AAV2/6 demonstrated the highest transduction efficiency in 3T3-L1 preadipocytes, as it was 1.5–2-fold more effective than AAV2/5, and AAV2/8 in the range of viral concentrations from 2×104 to 1,6×105 VG/cell. AAV2/5 and AAV2/8 showed transduction effi-ciencies similar to each other. The expression of GFP under the CMV promoter remained stable for up to 20 days. The induction of 3T3-L1 differentiation in three days after AAV transduction did not alter the GFP expression level, and AAV2/6 showed the best transduction efficiency. AAV2/6 demonstrated the ability to transduce mature adipocytes. These results were confirmed by in vivo studies on C57BL6 mice AAV2/6 had the highest transducing activity on both inguinal and inter-scapular adipose tissue. Thus, AAV2/6 has demonstrated higher transduction efficacy compared to AAV2/5, AAV2/8 and AAV2/9 both in 3T3-L1 adipocytes and adipose tissue in vivo, which proves its usability along with AAV8 and AAV9 for gene delivery to adipocytes.
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
