CRISPR and Genome Editing – Multimedia

In this webinar, we will present experimental approaches and real-life examples for creating transgenic animal models using CRISPR.

Chemical synthesis has been easily applied for rapidly generating either crRNA and tracrRNA or synthetic sgRNA for direct delivery into cells for gene editing applications such as DNA-free options and high throughput arrayed screening.

CRISPR is revolutionizing the life sciences. Genome editing using RNA-programmable nucleases opens up a wealth of experimental options. At the same time, it makes creating new cell and animal models faster, easier and cheaper than in the past. Consequently, researchers in areas as diverse as cell/developmental biology, genetics or clinical research are turning to CRISPR as their method of choice.

Loss-of-function genetic screens with pooled sgRNA and shRNA libraries aid in discovery / prioritization of drug targets, understanding of disease progression and gene-disease associations, analysis of signal transduction pathways, characterization of the mechanisms for compounds, uncovering of synergistic gene interactions, and identification of markers of drug resistance and sensitivity. This webinar presents a comparison of loss-of-function genetic screens performed with CRISPR versus RNAi technology.

Precise genome engineering with CRISPR-Cas9 and single-stranded DNA oligo or double-stranded DNA plasmid donors via homology-directed repair (HDR).

We have developed protocols using our StemRNA-NM Reprogramming Kit for generating iPS cells from skin-derived fibroblasts, blood-derived EPCs, and urine-derived UDCs using GMP-compatible reagents (NutriStem medium, iMatrix-511 recombinant laminin). These protocols generate GMP-compatible stem cells without the use of feeders, conditioned medium, additional small molecules or proteins, or passaging during reprogramming.

Synthetic, dual-RNA-encoded Cas9 is used for precise homology-directed repair (HDR) gene engineering. Both short and long (GFP) inserts are covered.

CRISPR-Cas9 gene editing using synthetic crRNA:tracrRNA or sgRNA is highly efficient and easy to use. Synthetic crRNA:tracrRNA is uniquely suited to in vitro and in vivo applications, in particular, DNA-free approach with Cas9 mRNA. Chemical synthesis of guide RNAs allows accurate and rapid production of arrayed crRNA libraries for high-confidence, loss-of-function screens.

Joseph Wu, Director, Stanford Cardiovascular Institute, speaking at Stem Cells in Drug Discovery.

In the current study we have used siRNA targeting VEGFR2 as an example to study knockdown of VEGFR2 and subsequent inhibition of tube formation by HUVECs on Growth Factor Reduced Matrigel™ in a 96-well plate format. The same strategy can be used for screening and validating siRNA based inhibitors of the angiogenic process in vitro and thus could be of utility in anti-cancer screening strategies.