Establishing the allosteric mechanism in CRISPR‐Cas9 - Publikacja - MOST Wiedzy

Wyszukiwarka

Establishing the allosteric mechanism in CRISPR‐Cas9

Abstrakt

Allostery is a fundamental property of proteins, which regulates biochemical information transfer between spatially distant sites. Here, we report on the critical role of molecular dynamics (MD) simulations in discovering the mechanism of allosteric communication within CRISPR‐Cas9, a leading genome editing machinery with enormous promises for medicine and biotechnology. MD revealed how allostery intervenes during at least three steps of the CRISPR‐Cas9 function: affecting DNA recognition, mediating the cleavage and interfering with the off‐target activity. An allosteric communication that activates concerted DNA cleavages was found to led through the L1/L2 loops, which connect the HNH and RuvC catalytic domains. The identification of these “allosteric transducers” inspired the development of novel variants of the Cas9 protein with improved specificity, opening a new avenue for controlling the CRISPR‐Cas9 activity. Discussed studies also highlight the critical role of the recognition lobe in the conformational activation of the catalytic HNH domain. Specifically, the REC3 region was found to modulate the dynamics of HNH by sensing the formation of the RNA:DNA hybrid. The role of REC3 was revealed to be particularly relevant in the presence of DNA mismatches. Indeed, interference of REC3 with the RNA:DNA hybrid containing mismatched pairs at specific positions resulted in locking HNH in an inactive “conformational checkpoint” conformation, thereby hampering off‐target cleavages. Overall, MD simulations established the fundamental mechanisms underlying the allosterism of CRISPR‐Cas9, aiding engineering strategies to develop new CRISPR‐Cas9 variants for improved genome editing.

Cytowania

  • 3 3

    CrossRef

  • 0

    Web of Science

  • 3 3

    Scopus

Autorzy (4)

Cytuj jako

Pełna treść

pobierz publikację
pobrano 27 razy
Wersja publikacji
Accepted albo Published Version
Licencja
Copyright (2020 Wiley Periodicals LLC)

Słowa kluczowe

Informacje szczegółowe

Kategoria:
Publikacja w czasopiśmie
Typ:
artykuły w czasopismach
Opublikowano w:
Wiley Interdisciplinary Reviews-Computational Molecular Science nr 11,
ISSN: 1759-0876
Język:
angielski
Rok wydania:
2021
Opis bibliograficzny:
Nierzwicki Ł., Arantes P., Saha A., Palermo G.: Establishing the allosteric mechanism in CRISPR‐Cas9// Wiley Interdisciplinary Reviews-Computational Molecular Science -Vol. 11,iss. 3 (2021), s.e1503-
DOI:
Cyfrowy identyfikator dokumentu elektronicznego (otwiera się w nowej karcie) 10.1002/wcms.1503
Weryfikacja:
Politechnika Gdańska

wyświetlono 66 razy

Publikacje, które mogą cię zainteresować

Meta Tagi