DIPA-CRISPR is a simple and accessible method for insect gene editing

Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for injection. To overcome thes...

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Autores: Shirai, Yu, Piulachs, Maria-Dolors, Bellés, Xavier, Daimon, Takaaki
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/272293
Acceso en línea:http://hdl.handle.net/10261/272293
Access Level:acceso abierto
Palabra clave:CRISPR-Cas9
DIPA-CRISPR
Gene editing
Genome editing
Cockroaches
Beetles
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spelling DIPA-CRISPR is a simple and accessible method for insect gene editing Shirai, Yu Piulachs, Maria-Dolors Bellés, Xavier Daimon, Takaaki CRISPR-Cas9 DIPA-CRISPR Gene editing Genome editing Cockroaches Beetles Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for injection. To overcome these limitations, we report a simple and accessible method for insect gene editing, termed “direct parental” CRISPR (DIPA-CRISPR). We show that injection of Cas9 ribonucleoproteins (RNPs) into the haemocoel of adult females efficiently introduces heritable mutations in developing oocytes. Importantly, commercially available standard Cas9 protein can be directly used for DIPA-CRISPR, which makes this approach highly practical and feasible. DIPA-CRISPR enables highly efficient gene editing in the cockroaches, on which conventional approaches cannot be applied, and in the model beetle Tribolium castaneum. Due to its simplicity and accessibility, DIPA-CRISPR will greatly extend the application of gene editing technology to a wide variety of insects. This research was supported by JSPS KAKENHI (nos. 20H02999 and 20K21311 to T.D.); JSPS Open Partnership Joint Research Projects (no. JPJSBP120209917 to T.D.); Spanish Ministry of Innovation and Competitiveness (PID2019-104483GB-I00 to X.B.); CSIC-Spain (2019AEP028 to M.-D.P.); and in part by Cabinet Office, Government of Japan, Cross-ministerial Moonshot Agriculture, Forestry and Fisheries Research and Development Program (no. JPJ009237 to T.D.). Y.S. is a JSPS Research Fellow with a research grant (DC1, 21J20658). Peer reviewed Elsevier http://hdl.handle.net/10261/272293
title DIPA-CRISPR is a simple and accessible method for insect gene editing
spellingShingle DIPA-CRISPR is a simple and accessible method for insect gene editing
Shirai, Yu
CRISPR-Cas9
DIPA-CRISPR
Gene editing
Genome editing
Cockroaches
Beetles
title_short DIPA-CRISPR is a simple and accessible method for insect gene editing
title_full DIPA-CRISPR is a simple and accessible method for insect gene editing
title_fullStr DIPA-CRISPR is a simple and accessible method for insect gene editing
title_full_unstemmed DIPA-CRISPR is a simple and accessible method for insect gene editing
title_sort DIPA-CRISPR is a simple and accessible method for insect gene editing
author Shirai, Yu
author_facet Shirai, Yu
Piulachs, Maria-Dolors
Bellés, Xavier
Daimon, Takaaki
author_role author
author2 Piulachs, Maria-Dolors
Bellés, Xavier
Daimon, Takaaki
author2_role author
author
author
topic CRISPR-Cas9
DIPA-CRISPR
Gene editing
Genome editing
Cockroaches
Beetles
topic_facet CRISPR-Cas9
DIPA-CRISPR
Gene editing
Genome editing
Cockroaches
Beetles
description Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for injection. To overcome these limitations, we report a simple and accessible method for insect gene editing, termed “direct parental” CRISPR (DIPA-CRISPR). We show that injection of Cas9 ribonucleoproteins (RNPs) into the haemocoel of adult females efficiently introduces heritable mutations in developing oocytes. Importantly, commercially available standard Cas9 protein can be directly used for DIPA-CRISPR, which makes this approach highly practical and feasible. DIPA-CRISPR enables highly efficient gene editing in the cockroaches, on which conventional approaches cannot be applied, and in the model beetle Tribolium castaneum. Due to its simplicity and accessibility, DIPA-CRISPR will greatly extend the application of gene editing technology to a wide variety of insects.
publishDate 2022
format article
status_str publishedVersion
url http://hdl.handle.net/10261/272293
eu_rights_str_mv openAccess
publisher Elsevier
institution Consejo Superior de Investigaciones Científicas (CSIC)
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
_version_ 1878437221047992320
publishDateSort 2022
author_browse Bellés, Xavier
Daimon, Takaaki
Piulachs, Maria-Dolors
Shirai, Yu
publisherStr Elsevier
score 6,924472

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