Engineering Anti-CRISPR Proteins to Create CRISPR-Cas Protein Switches for Activatable Genome Editing and Viral Protease Detection
© 2024 Wiley‐VCH GmbH..
Proteins capable of switching between distinct active states in response to biochemical cues are ideal for sensing and controlling biological processes. Activatable CRISPR-Cas systems are significant in precise genetic manipulation and sensitive molecular diagnostics, yet directly controlling Cas protein function remains challenging. Herein, we explore anti-CRISPR (Acr) proteins as modules to create synthetic Cas protein switches (CasPSs) based on computational chemistry-directed rational protein interface engineering. Guided by molecular fingerprint analysis, electrostatic potential mapping, and binding free energy calculations, we rationally engineer the molecular interaction interface between Cas12a and its cognate Acr proteins (AcrVA4 and AcrVA5) to generate a series of orthogonal protease-responsive CasPSs. These CasPSs enable the conversion of specific proteolytic events into activation of Cas12a function with high switching ratios (up to 34.3-fold). These advancements enable specific proteolysis-inducible genome editing in mammalian cells and sensitive detection of viral protease activities during virus infection. This work provides a promising strategy for developing CRISPR-Cas tools for controllable gene manipulation and regulation and clinical diagnostics.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:63 |
|---|---|
| Enthalten in: |
Angewandte Chemie (International ed. in English) - 63(2024), 16 vom: 15. Apr., Seite e202400599 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Kang, Wenyuan [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Anti-CRISPR Protein |
|---|
| Anmerkungen: |
Date Completed 10.04.2024 Date Revised 10.04.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1002/anie.202400599 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM368975169 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM368975169 | ||
| 003 | DE-627 | ||
| 005 | 20240410232550.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 240229s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1002/anie.202400599 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1371.xml |
| 035 | |a (DE-627)NLM368975169 | ||
| 035 | |a (NLM)38407550 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Kang, Wenyuan |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Engineering Anti-CRISPR Proteins to Create CRISPR-Cas Protein Switches for Activatable Genome Editing and Viral Protease Detection |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 10.04.2024 | ||
| 500 | |a Date Revised 10.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2024 Wiley‐VCH GmbH. | ||
| 520 | |a Proteins capable of switching between distinct active states in response to biochemical cues are ideal for sensing and controlling biological processes. Activatable CRISPR-Cas systems are significant in precise genetic manipulation and sensitive molecular diagnostics, yet directly controlling Cas protein function remains challenging. Herein, we explore anti-CRISPR (Acr) proteins as modules to create synthetic Cas protein switches (CasPSs) based on computational chemistry-directed rational protein interface engineering. Guided by molecular fingerprint analysis, electrostatic potential mapping, and binding free energy calculations, we rationally engineer the molecular interaction interface between Cas12a and its cognate Acr proteins (AcrVA4 and AcrVA5) to generate a series of orthogonal protease-responsive CasPSs. These CasPSs enable the conversion of specific proteolytic events into activation of Cas12a function with high switching ratios (up to 34.3-fold). These advancements enable specific proteolysis-inducible genome editing in mammalian cells and sensitive detection of viral protease activities during virus infection. This work provides a promising strategy for developing CRISPR-Cas tools for controllable gene manipulation and regulation and clinical diagnostics | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Anti-CRISPR Protein | |
| 650 | 4 | |a CRISPR-Cas | |
| 650 | 4 | |a Protein Engineering | |
| 650 | 4 | |a Protein Switch | |
| 650 | 4 | |a Viral Protease Detection | |
| 650 | 7 | |a Peptide Hydrolases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 650 | 7 | |a CRISPR-Associated Proteins |2 NLM | |
| 650 | 7 | |a Endopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 650 | 7 | |a Viral Proteases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 700 | 1 | |a Xiao, Fei |e verfasserin |4 aut | |
| 700 | 1 | |a Zhu, Xi |e verfasserin |4 aut | |
| 700 | 1 | |a Ling, Xinyu |e verfasserin |4 aut | |
| 700 | 1 | |a Xie, Shiyi |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Ruimiao |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Peihang |e verfasserin |4 aut | |
| 700 | 1 | |a Cao, Linxin |e verfasserin |4 aut | |
| 700 | 1 | |a Lei, Chunyang |e verfasserin |4 aut | |
| 700 | 1 | |a Qiu, Ye |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Tao |e verfasserin |4 aut | |
| 700 | 1 | |a Nie, Zhou |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Angewandte Chemie (International ed. in English) |d 1964 |g 63(2024), 16 vom: 15. Apr., Seite e202400599 |w (DE-627)NLM000105422 |x 1521-3773 |7 nnns |
| 773 | 1 | 8 | |g volume:63 |g year:2024 |g number:16 |g day:15 |g month:04 |g pages:e202400599 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1002/anie.202400599 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 63 |j 2024 |e 16 |b 15 |c 04 |h e202400599 | ||