# Administrative Supplement for Direct and rapid control of proteins at scale

> **NIH NIH DP2** · CHILDREN'S HOSP OF PHILADELPHIA · 2022 · $156,648

## Abstract

Abstract:
Since the completion of the Human Genome Project, a major effort in the biomedical sciences has been to
understand the function of proteins using perturbation. Functional screening platforms based on RNAi and
CRISPR, in particular, have been instrumental in pursuing this goal. Yet despite their wide use and
extraordinary strengths, even todays state-of-the-art platforms are still limited in their capabilities. Two
prominent limitations include (1) The inability to study the function of genes that are essential for cellular
growth, and (2) the slow dynamics of protein depletion that enable adaptation mechanisms that can
influence the results. To overcome these limitations, I propose to develop a novel pooled screening
platform based on rapid and direct modulation of proteins in human cells at a genome-scale. This
platform will be based on multiplexed endogenous tagging of genes with HaloTag, a bioorthogonal protein
domain capable of binding a variety of small molecule ligands that can be used to either increase or
decrease the stability of the fusion protein. In this proposal I will optimize pooled multiplexed endogenous
gene tagging using a unique intron targeting approach which our lab has developed. Genome-scale
HaloTag fusion libraries will be used to screen for effects of acute protein degradation and destabilization,
identify genes that are prone for transcriptional adaptation, and identify aggregating proteins in an unbiased
manner using a different fluorescence ligands.

## Key facts

- **NIH application ID:** 10521748
- **Project number:** 3DP2GM137416-01S1
- **Recipient organization:** CHILDREN'S HOSP OF PHILADELPHIA
- **Principal Investigator:** Ophir Shalem
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $156,648
- **Award type:** 3
- **Project period:** 2021-12-01 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10521748

## Citation

> US National Institutes of Health, RePORTER application 10521748, Administrative Supplement for Direct and rapid control of proteins at scale (3DP2GM137416-01S1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10521748. Licensed CC0.

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