CRISPR/Cas9 genome editing introduction and optimization in the non-model insect Pyrrhocoris apterus

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F19%3A00506496" target="_blank" >RIV/60077344:_____/19:00506496 - isvavai.cz</a>

Alternative codes found

RIV/60076658:12310/19:43899510

Result on the web

<a href="https://www.frontiersin.org/articles/10.3389/fphys.2019.00891/full" target="_blank" >https://www.frontiersin.org/articles/10.3389/fphys.2019.00891/full</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3389/fphys.2019.00891" target="_blank" >10.3389/fphys.2019.00891</a>

Result language

angličtina

Original language name

CRISPR/Cas9 genome editing introduction and optimization in the non-model insect Pyrrhocoris apterus

Original language description

The CRISPR/Cas9 technique is widely used in experimentation with human cell lines as well as with other model systems, such as mice Mus musculus, zebrafish Danio reiro, and the fruit fly Drosophila melanogaster. However, publications describing the use of CRISPR/Cas9 for genome editing in non-model organisms, including non-model insects, are scarce. The introduction of this relatively new method presents many problems even for experienced researchers, especially with the lack of procedures to tackle issues concerning the efficiency of mutant generation. Here we present a protocol for efficient genome editing in the non-model insect species Pyrrhocoris apterus. We collected data from several independent trials that targeted several genes using the CRISPR/Cas9 system and determined that several crucial optimization steps led to a remarkably increased efficiency of mutant production. The main steps are as follows: the timing of embryo injection, the use of the heteroduplex mobility assay as a screening method, in vivo testing of sgRNA efficiency, and G0 germline mosaicism screening. The timing and the method of egg injections used here need to be optimized for other species, but other here-described optimization solutions can be applied immediately for genome editing in other insect species.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10608 - Biochemistry and molecular biology

Project

<a href="/en/project/GA15-23681S" target="_blank" >GA15-23681S: Neuroendocrine control of insect reproductive diapause and metamorphosis</a><br>

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2019

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Frontiers in physiology

ISSN

1664-042X

e-ISSN

—

Volume of the periodical

10

Issue of the periodical within the volume

JUL 15

Country of publishing house

CH - SWITZERLAND

Number of pages

15

Pages from-to

891

UT code for WoS article

000475487600001

EID of the result in the Scopus database

2-s2.0-85069944729