# Intracellular CRISPR gRNA assembly for massively multiplexed, one pot, (epi)genetic screening

> **NIH NIH R21** · NORTH CAROLINA STATE UNIVERSITY RALEIGH · 2020 · $198,360

## Abstract

Project Summary
It has been clear for over a decade that cancer is not a single disease and that this heterogeneity is a primary
barrier to the understanding of oncogenic mechanisms and treatments. Cancers vary epigenetically and
genetically at multiple length and time scales and between patients. There can be many distinct mechanisms
driving oncogenic and metastatic processes, even within a single cancerous cell. The challenge researchers and
clinicians face is how to understand cancer from the perspective of simultaneous perturbations to multiple
genes and proteins. Risk variants identified through genome wide association studies and epiGWAS can be
individually perturbed in large experiments comprised of many 384-well plates through RNAi and CRISPR
libraries, and even be combinatorially perturbed and screened in `one-pot' using barcoding strategies.
However, even state-of-the-art CRISPR screening methods are restricted to functional perturbations of 2 or 3
variants at a time per cell. These restrictions arise from the difficulties repetitive sequences in gRNA arrays
present in both expression construct synthesis and stability. Here we propose a new method that will be
capable of expressing randomized combinatorial libraries of thousands of distinct gRNAs, with each cell of a
population expressing an array of over 30 gRNAs.

## Key facts

- **NIH application ID:** 9999547
- **Project number:** 5R21CA240162-02
- **Recipient organization:** NORTH CAROLINA STATE UNIVERSITY RALEIGH
- **Principal Investigator:** Albert Keung
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $198,360
- **Award type:** 5
- **Project period:** 2019-09-01 → 2022-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9999547

## Citation

> US National Institutes of Health, RePORTER application 9999547, Intracellular CRISPR gRNA assembly for massively multiplexed, one pot, (epi)genetic screening (5R21CA240162-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9999547. Licensed CC0.

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