U.S. flag

An official website of the United States government

Format

Send to:

Choose Destination

SRX6721807: GSM4037149: Cas13d_DND_6h_554_Rep2; Danio rerio; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 37.3M spots, 2.8G bases, 908.7Mb downloads

Submitted by: NCBI (GEO)
Study: CRISPR-Cas13d induces efficient mRNA knock-down in animal embryos
show Abstracthide Abstract
Early embryonic development is driven by maternal gene products deposited into the oocyte. Due to the lack of reliable knockdown strategies, maternal mRNA functions have remained elusive. CRISPR-Cas13 systems have recently been employed to induce RNA degradation in yeast, plants and mammalian cell lines. However, no systematic study of the potential of Cas13 has been carried out in an animal model system. Here, we show that CRISPR-Cas13d is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that both zygotically-expressed and maternally-provided transcripts are efficiently targeted, resulting in an 85% average decrease in transcript level and the recapitulation of well-known embryonic phenotypes. Moreover, we show that this system can be used in medaka, killifish, sea anemone and mouse embryos. All together our results demonstrate that CRISPR-Cas13d is an efficient knockdown platform to understand gene function in animal embryos. Overall design: Zebrafish embryos were injected with the indicated mRNA and/or guideRNAs and after 6 hours post injection.There are two biological replicates per treatment.
Sample: Cas13d_DND_6h_554_Rep2
SAMN12591309 • SRS5284979 • All experiments • All runs
Organism: Danio rerio
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was extracted using Trizol mRNAseq libraries were generated from 500 ng of high-quality total RNA, as assessed using the Bioanalyzer (Agilent). Libraries were made according to the manufacturer's directions for the TruSeq Stranded mRNA LP Sample Prep Kit (Illumina, Cat. No. 20020594) with TruSeq RNA Single Indexes Set A and B (Illumina, Cat. No. 20020492 and 20020493). Resulting short fragment libraries were checked for quality and quantity using the Bioanalyzer (Agilent) and Qubit Fluorometer (Life Technologies). Libraries were pooled, re-quantified and sequenced as 75bp single reads on a high-output flowcell using the Illumina NextSeq instrument. Poly(A)_enrichment
Links:
Runs: 1 run, 37.3M spots, 2.8G bases, 908.7Mb
Run# of Spots# of BasesSizePublished
SRR997456537,283,9252.8G908.7Mb2022-01-23

ID:
8858916

Supplemental Content

Search details

See more...

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...