This is a Preprint and has not been peer reviewed. The published version of this Preprint is available: https://doi.org/10.1016/j.xpro.2023.102501. This is version 2 of this Preprint.
Two-step CRISPR-Cas9 protocol for transposable element deletion in D. melanogaster natural populations
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Abstract
This protocol generates a precise deletion of a transposable element (TE) in a natural population of Drosophila melanogaster using two steps of CRISPR-Cas9 homology-directed repair. In the first step, the TE is replaced by a fluorescent marker, while in the second CRISPR-Cas9 step the fluorescence marker is removed to avoid any possible effect of the introduced marker sequence. This two-step protocol thus produces a precise deletion of any genomic region, exemplified here with a TE, while facilitating the screening of positive CRISPR-Cas9 events in natural populations without altering their genetic background.
For complete details on the use and execution of this protocol, please refer to (Merenciano & Gonzalez, 2023).
DOI
https://doi.org/10.32942/X2P88M
Subjects
Life Sciences
Keywords
CRISPR/Cas9, Drosophila, natural population
Dates
Published: 2023-02-15 16:56
Last Updated: 2023-02-15 21:56
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License
CC-By Attribution-NonCommercial-NoDerivatives 4.0 International
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Conflict of interest statement:
None
Data and Code Availability Statement:
Not applicable
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