CRISPR-Mediated Cassette Exchange (CriMCE): A Method to Introduce and Isolate Precise Marker-Less Edits

Morianou, Ioanna, Crisanti, Andrea, Nolan, Tony ORCID: https://orcid.org/0000-0002-2982-8333 and Hammond, Andrew M (2022) 'CRISPR-Mediated Cassette Exchange (CriMCE): A Method to Introduce and Isolate Precise Marker-Less Edits'. The CRISPR Journal, Vol 5, Issue 6, pp. 868-876.

	Text submitted CriMCE accepted.pdf - Accepted Version Restricted to Repository staff only Download (4MB)
Preview	Text Morianou_ManuscriptRevised_withFigs.pdf - Accepted Version Download (1MB) | Preview

Abstract

The introduction of small unmarked edits to the genome of insects is essential to study the molecular underpinnings of important biological traits, such as resistance to insecticides and genetic control strategies. Advances in CRISPR genome engineering have made this possible, but prohibitively laborious for most laboratories due to low rates of editing and the lack of a selectable marker. To facilitate the generation and isolation of precise marker-less edits we have developed a two-step method based on CRISPR-mediated cassette exchange (CriMCE) of a marked placeholder for a variant of interest. This strategy can be used to introduce a wider range of potential edits compared with previous approaches while consolidating the workflow. We present proof-of-principle that CriMCE is a powerful tool by engineering three single nucleotide polymorphism variants into the genome of Anopheles gambiae, with 5–41 × higher rates of editing than homology-directed repair or prime editing.

Item Type:	Article
Additional Information:	This is the accepted version of the following article: Ioanna Morianou, Andrea Crisanti, Tony Nolan, and Andrew M. Hammond. CRISPR-Mediated Cassette Exchange (CriMCE): A Method to Introduce and Isolate Precise Marker-Less Edits. The CRISPR Journal.Dec 2022.868-876, which has now been formally published in final form at The Crispr Journal at http://doi.org/10.1089/crispr.2022.0026. This original submission version of the article may be used for non-commercial purposes in accordance with the Mary Ann Liebert, Inc., publishers’ self-archiving terms and conditions.
Subjects:	QU Biochemistry > Genetics > QU 460 Genomics. Proteomics QU Biochemistry > Genetics > QU 550 Genetic techniques. PCR. Chromosome mapping QX Parasitology > QX 20 Research (General) QX Parasitology > Insects. Other Parasites > QX 515 Anopheles
Faculty: Department:	Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI):	https://doi.org/10.1089/crispr.2022.0026
Depositing User:	Maria Grimes
Date Deposited:	01 Feb 2023 11:11
Last Modified:	04 Jul 2023 13:55
URI:	https://archive.lstmed.ac.uk/id/eprint/21745

Statistics

View details

Actions (login required)

Edit Item