Gene delivery to, and gene targeting in, stem cells would be a highly enabling technology for basic science and biomedical application. Adeno-associated viral (AAV) vectors have demonstrated the capacity for efficient delivery to numerous cells, but their application to stem cells has been limited by low transduction efficiency. Due to their considerable advantages, however, engineering AAV delivery systems to enhance gene delivery to stem cells may have an impact in stem cell biology and therapy. Therefore, using several diverse AAV capsid librariesincluding randomly mutagenized, DNA shuffled, and random peptide insertion variantswe applied directed evolution to create a designer AAV vector with enhanced delivery efficiency for neural stem cells (NSCs). A novel AAV variant, carrying an insertion of a selected peptide sequence on the surface of the threefold spike within the heparin-binding site, emerged from this evolution. Importantly, this evolved AAV variant mediated efficient gene delivery to rat, mouse, and human NSCs, as well as efficient gene targeting within adult NSCs, and it is thus promising for applications ranging from basic stem cell biology to clinical translation.
Bibliographical noteFunding Information:
This work was supported by California Institute for Regenerative Medicine Training grant number T1-00007 and grant number RT1-01021, and by National Research Foundation (NRF) grant funded by the Korea government (MEST) through the Active Polymer Center for Pattern Integration (No. R11-2007-050-00000-0). KIP was supported by Stem Cell Research Center grant and Healthcare Technology R&D Project grant funded by Korean Government. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of CIRM or any other agency of the State of California. The authors are also grateful to Prof Andrew Wurmser for kindly providing murine NSCs and Ilsun Kim for technical support on human NSC culture.
All Science Journal Classification (ASJC) codes
- Molecular Medicine
- Molecular Biology
- Drug Discovery