Computationally designed hyperactive Cas9 enzymes.

Animals CRISPR-Associated Protein 9 / genetics CRISPR-Cas Systems / genetics Gene Editing Genetic Engineering Genome Mammals

Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
31 05 2022
Historique:
received: 21 10 2021
accepted: 10 05 2022
entrez: 31 5 2022
pubmed: 1 6 2022
medline: 3 6 2022
Statut: epublish

Résumé

The ability to alter the genomes of living cells is key to understanding how genes influence the functions of organisms and will be critical to modify living systems for useful purposes. However, this promise has long been limited by the technical challenges involved in genetic engineering. Recent advances in gene editing have bypassed some of these challenges but they are still far from ideal. Here we use FuncLib to computationally design Cas9 enzymes with substantially higher donor-independent editing activities. We use genetic circuits linked to cell survival in yeast to quantify Cas9 activity and discover synergistic interactions between engineered regions. These hyperactive Cas9 variants function efficiently in mammalian cells and introduce larger and more diverse pools of insertions and deletions into targeted genomic regions, providing tools to enhance and expand the possible applications of CRISPR-based gene editing.

Identifiants

DOI: 10.1038/s41467-022-30598-9 PMID: 35641498 PMC: PMC9156780
pubmed: 35641498
doi: 10.1038/s41467-022-30598-9
pii: 10.1038/s41467-022-30598-9
pmc: PMC9156780
doi:

Substances chimiques

CRISPR-Associated Protein 9 EC 3.1.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

3023

Informations de copyright

© 2022. The Author(s).

Références

Proteins. 2015 Aug;83(8):1385-406
pubmed: 25670500
RNA. 2019 Jan;25(1):35-44
pubmed: 30348755
Nat Commun. 2020 Jul 17;11(1):3576
pubmed: 32681021
Nat Biotechnol. 2014 Apr;32(4):347-55
pubmed: 24584096
Biochem Soc Trans. 2013 Dec;41(6):1401-6
pubmed: 24256227
Mol Cell. 2019 Feb 7;73(3):611-620.e3
pubmed: 30606466
Nat Rev Microbiol. 2019 Jan;17(1):7-12
pubmed: 30171202
Nat Biotechnol. 2016 Mar;34(3):339-44
pubmed: 26789497
Yeast. 1998 Jan 30;14(2):115-32
pubmed: 9483801
Nucleic Acids Res. 2013 Apr;41(7):4336-43
pubmed: 23460208
Mol Cell. 2018 Jul 5;71(1):42-55.e8
pubmed: 29979968
Biotechniques. 2017 Jul 1;63(1):37-39
pubmed: 28701147
Science. 2018 Aug 31;361(6405):870-875
pubmed: 30166483
Bioinformatics. 2014 Aug 1;30(15):2114-20
pubmed: 24695404
Angew Chem Int Ed Engl. 2021 Mar 1;60(10):5554-5560
pubmed: 33300646
Science. 2018 Aug 31;361(6405):866-869
pubmed: 30166482
Nature. 2014 Mar 6;507(7490):62-7
pubmed: 24476820
Science. 2016 Feb 19;351(6275):867-71
pubmed: 26841432
J Am Chem Soc. 2018 Feb 28;140(8):2971-2984
pubmed: 29442507
Mol Cell. 2018 Oct 4;72(1):178-186.e5
pubmed: 30270109
Curr Opin Microbiol. 2017 Jun;37:88-94
pubmed: 28645099
Bioinformatics. 2011 Nov 1;27(21):2957-63
pubmed: 21903629
Mol Cell. 2018 Jun 7;70(5):801-813.e6
pubmed: 29804829
Cell Rep. 2018 Jan 9;22(2):359-371
pubmed: 29320733
Sci Adv. 2017 Aug 04;3(8):eaao0027
pubmed: 28808686
Methods Enzymol. 2002;350:87-96
pubmed: 12073338
Nat Methods. 2012 Mar 04;9(4):357-9
pubmed: 22388286
F1000Res. 2019 Nov 7;8:1874
pubmed: 33552473
Genome Res. 2002 Apr;12(4):656-64
pubmed: 11932250
Trends Biotechnol. 2019 Apr;37(4):389-401
pubmed: 30352704
Gastroenterology. 2002 Jan;122(1):211-9
pubmed: 11781295
Science. 2016 Jan 1;351(6268):84-8
pubmed: 26628643
Science. 2015 Jun 26;348(6242):1477-81
pubmed: 26113724
Cell. 2014 Jun 5;157(6):1262-1278
pubmed: 24906146
Nat Methods. 2015 Apr;12(4):357-60
pubmed: 25751142
Nature. 2022 Mar;603(7900):343-347
pubmed: 35236982

Auteurs

Pascal D Vos (PD)

Curtin Medical School, Curtin University, Bentley, WA, Australia.
Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
Centre for Medical Research, The University of Western Australia, Nedlands, WA, Australia.
School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia.
ORCID: 0000-0002-5770-8283

Giulia Rossetti (G)

Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
Telethon Kids Institute, Northern Entrance, Perth Children's Hospital, Nedlands, WA, Australia.

Jessica L Mantegna (JL)

Curtin Medical School, Curtin University, Bentley, WA, Australia.
Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.

Stefan J Siira (SJ)

Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
ORCID: 0000-0002-8391-0143

Andrianto P Gandadireja (AP)

Curtin Medical School, Curtin University, Bentley, WA, Australia.
Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
ORCID: 0000-0002-1847-2079

Mitchell Bruce (M)

Curtin Medical School, Curtin University, Bentley, WA, Australia.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.

Samuel A Raven (SA)

Curtin Medical School, Curtin University, Bentley, WA, Australia.
Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
Centre for Medical Research, The University of Western Australia, Nedlands, WA, Australia.

Olga Khersonsky (O)

Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.

Sarel J Fleishman (SJ)

Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.

Aleksandra Filipovska (A)

Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia.
Centre for Medical Research, The University of Western Australia, Nedlands, WA, Australia.
School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia.
Telethon Kids Institute, Northern Entrance, Perth Children's Hospital, Nedlands, WA, Australia.
ORCID: 0000-0002-6998-8403

Oliver Rackham (O)

Curtin Medical School, Curtin University, Bentley, WA, Australia. oliver.rackham@curtin.edu.au.
Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia. oliver.rackham@curtin.edu.au.
Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, WA, Australia. oliver.rackham@curtin.edu.au.
ARC Centre of Excellence in Synthetic Biology, QEII Medical Centre, Nedlands, WA, Australia. oliver.rackham@curtin.edu.au.
Telethon Kids Institute, Northern Entrance, Perth Children's Hospital, Nedlands, WA, Australia. oliver.rackham@curtin.edu.au.
ORCID: 0000-0002-5301-9624

Articles similaires

Evaluating the efficacy of telesurgery with dual console SSI Mantra Surgical Robotic System: experiment on animal model and clinical trials.

Sudhir Prem Srivastava, Vishwajyoti Pascual Srivastava, Avinesh Singh et al.
1.00
Robotic Surgical Procedures Animals Humans Telemedicine Models, Animal

Odour generalisation and detection dog training.

Lyn Caldicott, Thomas W Pike, Helen E Zulch et al.
1.00
Animals Odorants Dogs Generalization, Psychological Smell

FBXO22 inhibits colitis and colorectal carcinogenesis by regulating the degradation of the S2448-phosphorylated form of mTOR.

Minle Li, Xuan Chen, Pengfei Qu et al.
1.00
Animals TOR Serine-Threonine Kinases Colorectal Neoplasms Colitis Mice

Use of organic material provided by an automatic enrichment device by weaner pigs and its influence on tail lesions.

Philipp Heseker, Jeanette Probst, Stefanie Ammer et al.
1.00
Animals Tail Swine Behavior, Animal Animal Husbandry

Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines associées aux CRISPR › Protéine-9 associée à CRISPR › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Édition de gène › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Computationally designed hyperactive Cas9 enzymes.
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Computationally designed hyperactive Cas9 enzymes.