Vosshall Laboratory Aedes aegypti CRISPR‑Cas9 Genome‑Engineering Protocol

A step‑by‑step protocol for performing CRISPR‑Cas9 genome editing in the mosquito Aedes aegypti.


Vosshall Laboratory Aedes aegypti CRISPR‑Cas9 Genome‑Engineering Protocol - cover page
Brand
Vosshall Laboratory
Category
Other
Document type
Application Note
Language
English
Pages
17
File format
PDF
File size
599 KB
Published
30 September, 2026
Updated
30 September, 2026
MD5 checksum
3802AD0F6ACB672BE844F80E07F4DF14

About this manual

This protocol guide describes the complete workflow for generating precise genomic alterations in Aedes aegypti using the CRISPR‑Cas9 system. It covers target design, sgRNA synthesis, Cas9 source selection, donor DNA preparation, injection mix cleanup, microinjection, and downstream screening.

The document also lists recommended reagents, detailed reaction conditions, and troubleshooting tips, enabling researchers to reproduce the method in their own laboratories.

What's inside

  • Goals
  • General considerations
  • Selected Kits & Reagents
  • Target design and selection
  • sgRNA template generation and RNA production
  • In vitro transcription of sgRNAs
  • Choosing a source of Cas9
  • Donor plasmid or ssODN production for homology‑directed repair
  • Final clean‑up and injection
  • Screening for modification
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