Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d
In this study, the authors compared the efficiency of automated robotic and manual injection methods for the CRISPR-RfxCas13d (CasRx) system for mRNA knockdown and Cas9-mediated DNA targeting in zebrafish embryos. They targeted the no tail (TBXTA) gene as a proof-of-principle, evaluating the induced embryonic phenotypes. Both Cas9 and CasRx systems caused loss of function phenotypes for TBXTA. Cas9 protein exhibited a higher percentage of severe phenotypes compared with mRNA, while CasRx protein and mRNA showed similar efficiency. Both robotic and manual injections demonstrated comparable phenotype percentages and mortality rates. The findings highlight the potential of RNA-targeting CRISPR effectors for precise gene knockdown and endorse automated microinjection at a speed of 1.0 s per embryo as a high-throughput alternative to manual methods.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:76 |
|---|---|
| Enthalten in: |
BioTechniques - 76(2024), 5 vom: 15. Apr., Seite 183-191 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Del Prado, Joaquin Abugattas-Nuñez [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
CasRx |
|---|
| Anmerkungen: |
Date Completed 26.04.2024 Date Revised 26.04.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.2144/btn-2023-0062 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM369108558 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM369108558 | ||
| 003 | DE-627 | ||
| 005 | 20240427232120.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 240301s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.2144/btn-2023-0062 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1390.xml |
| 035 | |a (DE-627)NLM369108558 | ||
| 035 | |a (NLM)38420933 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Del Prado, Joaquin Abugattas-Nuñez |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Comparing robotic and manual injection methods in zebrafish embryos for high-throughput RNA silencing using CRISPR-RfxCas13d |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 26.04.2024 | ||
| 500 | |a Date Revised 26.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a In this study, the authors compared the efficiency of automated robotic and manual injection methods for the CRISPR-RfxCas13d (CasRx) system for mRNA knockdown and Cas9-mediated DNA targeting in zebrafish embryos. They targeted the no tail (TBXTA) gene as a proof-of-principle, evaluating the induced embryonic phenotypes. Both Cas9 and CasRx systems caused loss of function phenotypes for TBXTA. Cas9 protein exhibited a higher percentage of severe phenotypes compared with mRNA, while CasRx protein and mRNA showed similar efficiency. Both robotic and manual injections demonstrated comparable phenotype percentages and mortality rates. The findings highlight the potential of RNA-targeting CRISPR effectors for precise gene knockdown and endorse automated microinjection at a speed of 1.0 s per embryo as a high-throughput alternative to manual methods | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Comparative Study | |
| 650 | 4 | |a CasRx | |
| 650 | 4 | |a high throughput | |
| 650 | 4 | |a knockdown | |
| 650 | 4 | |a robot microinjector | |
| 650 | 4 | |a zebrafish | |
| 650 | 7 | |a Zebrafish Proteins |2 NLM | |
| 650 | 7 | |a RNA, Messenger |2 NLM | |
| 700 | 1 | |a Ding, Yi |e verfasserin |4 aut | |
| 700 | 1 | |a Sonneville, Jan de |e verfasserin |4 aut | |
| 700 | 1 | |a der Kolk, Kees-Jan van |e verfasserin |4 aut | |
| 700 | 1 | |a Moreno-Mateos, Miguel A |e verfasserin |4 aut | |
| 700 | 1 | |a Málaga-Trillo, Edward |e verfasserin |4 aut | |
| 700 | 1 | |a Spaink, Herman P |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t BioTechniques |d 1988 |g 76(2024), 5 vom: 15. Apr., Seite 183-191 |w (DE-627)NLM012627046 |x 1940-9818 |7 nnns |
| 773 | 1 | 8 | |g volume:76 |g year:2024 |g number:5 |g day:15 |g month:04 |g pages:183-191 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.2144/btn-2023-0062 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 76 |j 2024 |e 5 |b 15 |c 04 |h 183-191 | ||