Skip to main content

Identification and Isolation of Novel Sugar-Like RNA Protecting Materials: Glycylglycerins from Pluripotent Stem Cells

  • Protocol
  • First Online:
MicroRNA Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1733))

  • 1861 Accesses

Abstract

Pluripotent stem cells are a resourceful treasure box for regenerative medicine. They contain a large variety of novel materials useful for designing and developing new medicines and therapies directed against many aging-associated degenerative disorders, including Alzheimer’s disease, Parkinson’s disease, stroke, diabetes, osteoporosis, and cancers. Currently, identification of these novel stem cell-specific materials is one of major breakthroughs in the field of stem cell research. Particularly, since the discovery of induced pluripotent stem cells (iPSC) in year 2006, the methods of iPSC derivation further provide an unlimited resource for screening, isolating, and even producing theses novel stem cell-specific materials in vitro. Using iPSCs, we can now prepare high quality and quantity of pure stem cell-specific agents for testing their therapeutic functions in treating various illnesses. These newly found stem cell-specific agents are divided into four major categories, including proteins, saccharides, nucleic acids, and small molecules (chemicals). In this article, we herein disclose one of the methodologies for isolating and purifying glycylglycerins—a group of glycylated sugar alcohols that protect hairpin-like microRNA precursors (pre-miRNA) and some of tRNAs in pluripotent stem cells. In view of such a unique RNA-protecting feature, glycylglycerins may be used to preserve and deliver functional small RNAs, such as pre-miRNAs and small interfering RNAs (siRNA), into human cells for eliciting their specific RNA interference (RNAi) effects, which may greatly advance the use of RNAi technology for treating human diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 99.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 139.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Lin SL, Chang DC, Chang-Lin S, Lin CH, Wu DT, Chen DT, Ying SY (2008) Mir-302 reprograms human skin cancer cells into a pluripotent ES-cell-like state. RNA 14:2115–2124

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Lin SL, Chang DC, Lin CH, Ying SY, Leu D, Wu DT (2011) Regulation of somatic cell reprogramming through inducible mir-302 expression. Nucleic Acids Res 39:1054–1065

    Article  CAS  PubMed  Google Scholar 

  3. Lin SL, Ying SY (2012) Mechanism and method for generating tumor-free iPS cells using intronic microRNA miR302 induction. In: Ying SY (ed) MicroRNA Protocols, 2nd edn. Springer Publishers press, New York, pp 295–324

    Google Scholar 

  4. Chang-Lin S, Hung A, Chang DC, Lin YW, Ying SY, Lin SL (2016) Novel glycylated sugar alcohols protect ESC-specific microRNAs from degradation in iPS cells. Nucleic Acids Res 44:4894–4906

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Chen SKJ, Lin SL (2013) Recent patents on microRNA-induced pluripotent stem cell generation. Recent Pat Regen Med 3:5–16

    Google Scholar 

  6. Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663–676

    Article  CAS  PubMed  Google Scholar 

  7. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920

    Article  CAS  PubMed  Google Scholar 

  8. Wernig M, Meissner A, Foreman R, Brambrink T, Ku M, Hochedlinger K, Bernstein BE, Jaenisch R (2007) In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448:318–324

    Article  CAS  PubMed  Google Scholar 

  9. Li HH, Lin SL, Huang CN, Lu FJ, Chiu PY, Huang WN, Lai TJ, Lin CL (2016) miR-302 attenuates amyloid-β-induced neurotoxicity through activation of Akt signaling. J Alzheimers Dis 50:1083–1098

    Article  CAS  PubMed  Google Scholar 

  10. Lin SL, Chang-Lin S, Lin YW, Chang D (2014) Use of novel monosaccharide-like glycylated sugar alcohol compositions for designing and developing anti-diabetic drugs. US patent application number 14/585978

    Google Scholar 

  11. Lin SL, Chang D, Ying SY, Leu D, Wu DTS (2010) MicroRNA miR-302 inhibits the tumorigenecity of human pluripotent stem cells by coordinate suppression of CDK2 and CDK4/6 cell cycle pathways. Cancer Res 70:9473–9482

    Article  CAS  PubMed  Google Scholar 

  12. Lin SL, Lin YW (2016) Sugar alcohol-based compositions for delivering nucleic acid-based drugs in vivo and in vitro. US patent number 9387251

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Shi-Lung Lin .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lin, SL. (2018). Identification and Isolation of Novel Sugar-Like RNA Protecting Materials: Glycylglycerins from Pluripotent Stem Cells. In: Ying, SY. (eds) MicroRNA Protocols . Methods in Molecular Biology, vol 1733. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7601-0_24

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7601-0_24

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7600-3

  • Online ISBN: 978-1-4939-7601-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics