Optimizing CRISPR/Cas9-based gene manipulation in echinoderms.
Cas9
Microinjection
Morpholino antisense oligonucleotides (MASO)
Sea star
Sea urchin
Synthetic single guide RNAs
Journal
Developmental biology
ISSN: 1095-564X
Titre abrégé: Dev Biol
Pays: United States
ID NLM: 0372762
Informations de publication
Date de publication:
10 2022
10 2022
Historique:
received:
05
05
2022
revised:
11
07
2022
accepted:
14
07
2022
pubmed:
3
8
2022
medline:
14
9
2022
entrez:
2
8
2022
Statut:
ppublish
Résumé
The impact of new technology can be appreciated by how broadly it is used. Investigators that previously relied only on pharmacological approaches or the use of morpholino antisense oligonucleotide (MASO) technologies are now able to apply CRISPR-Cas9 to study biological problems in their model organism of choice much more effectively. The transitions to new CRISPR-based approaches could be enhanced, first, by standardized protocols and education in their applications. Here we summarize our results for optimizing the CRISPR-Cas9 technology in a sea urchin and a sea star, and provide advice on how to set up CRISPR-Cas9 experiments and interpret the results in echinoderms. Our goal through these protocols and sharing examples of success by other labs is to lower the activation barrier so that more laboratories can apply CRISPR-Cas9 technologies in these important animals.
Identifiants
pubmed: 35917936
pii: S0012-1606(22)00142-7
doi: 10.1016/j.ydbio.2022.07.008
pii:
doi:
Substances chimiques
Morpholinos
0
RNA, Guide
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
117-124Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM140897
Pays : United States
Informations de copyright
Copyright © 2022 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest Synthego, IDT, and Aldevron each provided free samples of Cas9 and sgRNAs to initiate this project. All products thereafter were purchased at market value.