NOV 15, 2018 08:00 AM PST

Precise Gene Editing of Human Pluripotent Stem Cells

Speakers
  • Assistant Professor, Department of Biomedical Engineering, Wisconsin Institute for Discovery, University of Wisconsin-Madison
    Biography
      Krishanu Saha is an Assistant Professor in the Department of Biomedical Engineering at the University of Wisconsin-Madison. He is also a member of the Wisconsin Institute for Discovery, Carbone Cancer Center, and Stem Cell and Regenerative Medicine Center as well as the National Academies' Forum on Regenerative Medicine. Prior to his arrival in Madison, Dr. Saha studied Chemical Engineering at Cornell University and at the University of California in Berkeley. He was a Society in Science: Branco-Weiss fellow at the Whitehead Institute for Biomedical Research at MIT and in the Science and Technology Studies program at Harvard University. Major thrusts of his lab involve gene editing and cell engineering of human cells found in the retina, central nervous system and blood.

    Abstract:

    CRISPR ribonucleoproteins (RNPs) can generate programmable gene edits, however imprecise editing and efficient delivery to human stem cells are key challenges. Here we describe novel biochemical techniques to assemble various biomolecules and coatings with nanoscale precision around a RNP. First, by modifying the sgRNA with a short S1m RNA aptamer, we developed a modular strategy, termed an “S1mplex,” to assemble Cas9 RNPs with biotinylated moieties. Using S1mplexes with biotinylated short oligonucleotides improves the ratio of precise to imprecise editing up to 18-fold over conventional methods approaching a ratio of 4 precise edits to every imprecise mutation, while assembly with fluorescent molecules allows selection and enrichment for cells with multiplexed gene edits. Second, we developed synthetic coatings for nonviral delivery of RNPs to human pluripotent stem cells. Combined, these strategies, which utilize chemically-defined, off-the-shelf reagents, have significant promise for gene editing applications in vitro (e.g., drug discovery, disease modeling) with human stem cells.


    Show Resources
    You May Also Like
    MAY 22, 2018 08:00 AM PDT
    C.E. CREDITS
    MAY 22, 2018 08:00 AM PDT
    DATE: May 22, 2018TIME: 08:00AM PDT The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that...
    MAY 03, 2018 11:00 AM PDT
    MAY 03, 2018 11:00 AM PDT
    DATE: May 3, 2018TIME: 11:00AM PDT, 2:00PM EDTWhile stress is one of the leading causes of neuropsychiatric disorders, the molecular underpinnings of how stress induces alterations in b...
    APR 27, 2018 10:00 AM PDT
    C.E. CREDITS
    APR 27, 2018 10:00 AM PDT
    DATE: April 27, 2018TIME: 10:00am PST, 1:00pm ESTGlioblastoma (GBM) and Medulloblastoma (MB) are the most common adult and paediatric brain tumours, both of which can have devastating c...
    AUG 15, 2018 08:00 AM PDT
    C.E. CREDITS
    AUG 15, 2018 08:00 AM PDT
    DATE: August 15, 2018TIME: 08:00AM PDT, 11:00AM EDTThe failure of current chemotherapeutic strategies in the fight against cancer can be largely attributed to the occurrence of drug res...
    JUN 29, 2018 09:00 AM PDT
    C.E. CREDITS
    JUN 29, 2018 09:00 AM PDT
    DATE: June 29, 2018TIME: 09:00AM PDT, 12:00PM EDT There is significant epidemiological evidence to suggest that the consumption of a high-broccoli diet is associated with a r...
    MAY 24, 2018 09:30 AM PDT
    C.E. CREDITS
    MAY 24, 2018 09:30 AM PDT
    DATE: May 24, 2018 TIME: 9:30PM PDT The current gold standard in in vitro pre-clinical cancer treatment screening remain cell lines,...
    Loading Comments...