CRISPR screen and single-cell sequencing project
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The goal of this project is to screen H9-GFP tagged hESCs with 4 curated libraries and then use single-cell sequencing to determine which gRNA was incorporated into which cell and gene expression in each cell. The four libraries and some annotation info is listed in this file (File:Crispr screen single cell sequencing library.xlsx).
Library Design:
- From the curated gene lists, take all spacers for each gene from the gRNA list.
- Check each spacer for multi-mapping and that each spacer maps to the correct location.
- Keep only the top 3 spacers for each gene.
- Include 3 negative controls in each library, barcode using bc_25mer barcodes.
- Order gChip. See 05/05/2015 Library Order.
- Use CRISPR Library Prep Protocol to prepare library.