# Precision targeting of bladder cancer using CRISPR

> **NIH NIH R01** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2024 · $566,310

## Abstract

ABSTRACT
Chromosome rearrangements (structural variants) are common in cancer and they drive tumor
development by altering the expression or function of oncogenes and tumor suppressor genes
by copy number alterations, formation of oncogenic fusions or by alterations of DNA elements
responsible for regulating cancer genes. We have developed an innovative CRISPR-based
approach to specifically target structural variant junctions (SVJs) in cancer cells without harming
normal cells. This approach, called KLIPP, is based on the use of a “split” enzyme approach
consisting of inactivated dCas9 fused to the endonuclease Fok1 (Fok1-dCas9). To activate the
Fok1 endonuclease, two Fok1-dCas9 complexes are brought together at the SVJ using SVJ-
targeting guide RNAs leading to the induction of toxic DNA double strand breaks. We have
obtained strong proof-of-concept for this approach in bladder cancer and other cancer cell
systems both in cell cultures and in vivo. In this proposal we will perform pre-clinical efficacy
testing of the Precision KLIPP Therapy approach in an orthotopic mouse model of bladder
cancer. The premise for targeting bladder cancer with this approach is first, locally invasive
bladder cancer is a common and deadly disease with few impactful therapeutic options.
Second, bladder cancers typically show genomic instability with many hundreds of SVJs that
could be targeted with our approach. Third, the bladder is ideal for local inter-vesicle delivery of
the CRISPR reagents because of the ability to achieve high local concentrations and avoiding
systemic effects. We will in this application pursue three specific aims: Aim #1: CRISPR screen
to map potential Fok1-dCas9 binding toxicity genome-wide. Aim #2: Delivery of CRISPR
reagents to bladder cells in culture. Aim #3: Assessment of efficacy of Precision KLIPP Therapy
in a pre-clinical HGSC mouse model. With this R01 funding, we will be able to advance this
project to the point of planning for human clinical trials.

## Key facts

- **NIH application ID:** 10776828
- **Project number:** 1R01CA285730-01
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Mats Ljungman
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $566,310
- **Award type:** 1
- **Project period:** 2024-02-09 → 2029-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10776828

## Citation

> US National Institutes of Health, RePORTER application 10776828, Precision targeting of bladder cancer using CRISPR (1R01CA285730-01). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10776828. Licensed CC0.

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