Name: Genoombewerking in de gelekoortsmug Aedes aegypti met behulp van CRISPR-Cas9 (Video) | JoVE Journal
Uploaded: 2025-03-21T13:40:00.000+00:00
Duration: 6 min 47 s
Description: Leer genoombewerking in Aedes aegypti met CRISPR-Cas9 voor het creëren van genetisch gemodificeerde muggenlijnen. Deze methode ondersteunt onderzoek naar populatieonderdrukking en gecontroleerde genexpressie.

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</ol>

<table><tbody><tr><td>10x Cas9 reactiebuffer</td><td>PNA Bio&nbsp;</td><td>CB01</td><td></td></tr><tr><td>Benchling software</td><td>Benchling</td><td>N/A</td><td>www.benchling.com</td></tr><tr><td>Cas9 verdunningsbuffer</td><td>PNA Bio&nbsp;</td><td>CB03</td><td></td></tr><tr><td>Cas9-eiwit</td><td>PNA Bio&nbsp;</td><td>CP01-50</td><td></td></tr><tr><td>DH5&alpha; E. coli competente cellen</td><td>New England Biolabs</td><td>C2987</td><td></td></tr><tr><td>Dubbelzijdig plakband</td><td>Scotch Permanent</td><td>3136</td><td></td></tr><tr><td>Drosophila-buisjes</td><td>Genesee Scientific&nbsp;</td><td>32-109</td><td></td></tr><tr><td>Filterpapieren&nbsp;</td><td>GE Healthcare Life Science&nbsp;</td><td>1450-042</td><td></td></tr><tr><td>Visvoer&nbsp;</td><td>Tetra</td><td>B00025Z6YI</td><td>goudvissenvlokken&nbsp;</td></tr><tr><td>Flugs</td><td>Genesee Scientific&nbsp;</td><td>AS273</td><td></td></tr><tr><td>Fluorescentiemicroscoop</td><td>Leica Microsystems&nbsp;</td><td>&nbsp;M165 FC</td><td></td></tr><tr><td>Genfragment</td><td>Integrated DNA Technologies</td><td>N/A</td><td></td></tr><tr><td>gRNA</td><td>Synthego</td><td>N/A</td><td></td></tr><tr><td>Halocarbon olie 700&nbsp;</td><td>Sigma-Aldrich</td><td>H8898</td><td></td></tr><tr><td>Injectiemicroscoop</td><td>Leica Microsystems&nbsp;</td><td>DM2000</td><td></td></tr><tr><td>JM109 &nbsp;E. coli competente cellen&nbsp;</td><td>Zymo Research</td><td>T3005</td><td></td></tr><tr><td>Microinjector</td><td>Eppendorf</td><td></td><td>FemtoJet 4x&nbsp;</td></tr><tr><td>Microloader tips voor het vullen van Femtotips&nbsp;</td><td>Eppendorf</td><td>E5242956003</td><td></td></tr><tr><td>Micromanipulator&nbsp;</td><td>Eppendorf</td><td></td><td>TransferMan 4r&nbsp;</td></tr><tr><td>Micropipettrekkers&nbsp;</td><td>Sutter Instrument&nbsp;</td><td>P-2000</td><td></td></tr><tr><td>Microscoopdekglas&nbsp;</td><td>Fisherbrand&nbsp;</td><td>12-542-B</td><td></td></tr><tr><td>Microscoopglasplaat&nbsp;</td><td>Eisco</td><td>12-550-A3</td><td></td></tr><tr><td>Muisbloed (levende muizen gebruikt voor voeding)</td><td>University of California</td><td>IACUC, Animal Use Protocol #S17187</td><td>Gebaten voor muggenbloedvoeding; details voldoen aan protocollen voor dierenetiek</td></tr><tr><td>NEB Q5 High-Fidelity DNA-polymerase&nbsp;</td><td>New England Biolabs</td><td>M0491S</td><td></td></tr><tr><td>PCR-zuiveringskit</td><td>Qiagen</td><td>28004</td><td></td></tr><tr><td>Plasmid Miniprep Kit&nbsp;</td><td>Zymo Research</td><td>D4036</td><td></td></tr><tr><td>Kwartsfilament&nbsp;</td><td>Sutter Instruments</td><td>QF100-70-10</td><td></td></tr><tr><td>Transcriptie Clean-Up Kit</td><td>Fisher Scientific</td><td>AM1908</td><td></td></tr><tr><td>Ultra-zuiver water&nbsp;</td><td>Life Technologies</td><td>10977-023</td><td></td></tr></tbody></table>

genoombewerking in de gelekoortsmug aedes aegypti met behulp van crispr-cas9

Genome Editing in the Yellow Fever Mosquito Aedes aegypti using CRISPR-Cas9

Genoombewerking in de gelekoortsmug Aedes aegypti met behulp van CRISPR-Cas9

The emergence of the clustered, regularly interspersed, short palindromic repeats (CRISPR)-Cas9 technology has revolutionized the genetic engineering ...

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Research

JoVE Journal

Genetics

Genetica

In this video, we demonstrate the delivery of gene drive construct into the embryo of Anopheles gambiae mosquito through embryo microinjection technique. This procedure allows for integration of  delivered DNA construct, through homologous recombination, into the germline cells to generate transformed mosquito population.

Mosquito Embryo Microinjection: A Technique to Deliver Exogenous DNA into Embryo Yolk of Anopheles Gambiae to Generate Germline Mutations

Beschrijven we een methode voor het genereren van glmS-op basis van voorwaardelijke vechtpartij mutanten in Plasmodium falciparum met behulp van CRISPR/Cas9 genoom bewerken.

CRISPR/Cas9 Gene bewerken te maken voorwaardelijke mutanten van menselijke malariaparasiet P. falciparum

Dit protocol beschrijft micro-injectieprocedures voor Culex quinquefasciatus embryo's die zijn geoptimaliseerd om te werken met CRISPR/Cas9 genbewerkingstools. Deze techniek kan efficiënt site-specifieke, erfelijke, kiembaanmutaties genereren die kunnen worden gebruikt voor het bouwen van genetische technologieën in deze onderbelichte ziektevector.

Embryo micro-injection technieken voor efficiënte site-specifieke mutagenese in Culex quinquefasciatus

CRISPR/Cas9 wordt steeds vaker gebruikt om genfunctie in niet-modelorganismen te karakteriseren. Dit protocol beschrijft hoe knock-out lijnen van Culex pipienste genereren, van het bereiden van injectiemengsels tot het verkrijgen en injecteren van muggenembryo's, evenals hoe geïnjecteerde muggen en hun nakomelingen op te wekken, te kruisen en te screenen op gewenste mutaties.

Embryo's van Culex-muggen voorbereiden en injecteren om null-mutaties te genereren met CRISPR/Cas9

Dit protocol beschrijft de stappen van CRISPR/Cas9 gerichte mutagenese bij zandvliegen: embryoverzameling, injectie, insectenopfok en identificatie, evenals selectie van mutaties van belang.

Zandvlieg (Phlebotomus papatasi) Embryo Microinjection voor CRISPR/Cas9 Mutagenese

Hier wordt een protocol gepresenteerd van Helicoverpa armigera (Hübner) embryo micro-injectie en knock-out mutant identificatie gemaakt door CRISPR / Cas9 genoombewerking. Gemuteerde insecten maken verder onderzoek naar genfunctie en interactie tussen verschillende genen in vivo mogelijk.

Embryo Microinjectie en Knockout Mutant Identificatie van CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)

Het protocol beschrijft hoe locatiegerichte modificaties in het genoom van Anopheles malariamuggen kunnen worden bereikt met behulp van het φC31-systeem . De beschreven modificaties omvatten zowel de integratie als de uitwisseling van transgene cassettes in het genoom van attP-dragende koppelingslijnen.

Site-Directed φC31-gemedieerde integratie en cassette-uitwisseling in Anopheles Vectors of Malaria

Dit artikel beschrijft een protocol voor snelle identificatie van indels geïnduceerd door CRISPR /Cas9 en selectie van mutante lijnen in de mug Aedes aegypti met behulp van hoge resolutie smeltanalyse.

Indel Detection na CRISPR/Cas9 Mutagenese met behulp van Hoge resolutie Melt Analysis in de Mosquito Aedes aegypti

Dit artikel presenteert de stapsgewijze protocollen voor CRISPR/Cas9-mutagenese van de Oosterse fruitvlieg Bactrocera dorsalis. Gedetailleerde stappen in dit gestandaardiseerde protocol zullen dienen als een nuttige gids voor het genereren van gemuteerde vliegen voor functionele genstudies in B. dorsalis.

Protocollen voor CRISPR/Cas9 Mutagenese van de Oosterse Fruitvlieg Bactrocera dorsalis

Het huidige protocol beschrijft hoe gemeenschappelijke levensparametergegevens bij Aedes aegypti-muggen kunnen worden gemeten, waaronder vruchtbaarheid, vleugelgrootte, vruchtbaarheid, geslachtsverhouding, levensvatbaarheid, ontwikkelingstijden, mannelijke bijdrage en levensduur van volwassenen. Deze metingen kunnen worden gebruikt om de fitheid van transgene muggen te beoordelen.

Kwantificering van fitnesskosten bij transgene Aedes aegypti-muggen

Leer genoombewerking in Aedes aegypti met CRISPR-Cas9 voor het creëren van genetisch gemodificeerde muggenlijnen. Deze methode ondersteunt onderzoek naar populatieonderdrukking en gecontroleerde genexpressie.

Genoombewerking in de gelekoortsmug Aedes aegypti met behulp van CRISPR-Cas9

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