Evidence map›Paper›PMID 33688075›Full record

ArticleDevelopment (Cambridge, England)2021

A single-plasmid approach for genome editing coupled with long-term lineage analysis in chick embryos.

Shashank Gandhi, Yuwei Li, Weiyi Tang, Jens B Christensen, Hugo A Urrutia, Felipe M Vieceli, Michael L Piacentino, Marianne E Bronner

Erratum issuedOpen access · hybridAbstract read
In one paragraph

Article in Development (Cambridge, England), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
0.8field-weighted citation impact, top 30% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

8 citing papers in PubMed, 11 citations in OpenAlex.

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4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 2 countries.

Shashank GandhiDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Yuwei LiDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Weiyi TangDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Jens B ChristensenDepartment of Neuroscience, University of Copenhagen, Copenhagen, Blegdamsvej 3B, 2200 Copenhagen N, Denmark.
Hugo A UrrutiaDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Felipe M VieceliDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Michael L PiacentinoDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Marianne E BronnerDivision of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
California Institute of Technology · USUniversity of Copenhagen · DK

Funding

Clonal analysis of the cranial neural crestR01DE027568 · NIDCR · CALIFORNIA INSTITUTE OF TECHNOLOGY · PI BRONNER, MARIANNE · 2018 to 2022
$2.2M
Characterizing the neural crest response to BMP signaling through gastrulation and neurulationK99DE029240 · NIDCR · CALIFORNIA INSTITUTE OF TECHNOLOGY · PI PIACENTINO, MICHAEL LOUIS · 2019 to 2021
$397k
NIDCR NIH HHS K99 DE029240NIDCR NIH HHS R01 DE027568
6 · The paper itself

Abstract

An important strategy for establishing mechanisms of gene function during development is through mutation of individual genes and analysis of subsequent effects on cell behavior. Here, we present a single-plasmid approach for genome editing in chick embryos to study experimentally perturbed cells in an otherwise normal embryonic environment. To achieve this, we have engineered a plasmid that encodes Cas9 protein, gene-specific guide RNA (gRNA), and a fluorescent marker within the same construct. Using transfection- and electroporation-based approaches, we show that this construct can be used to perturb gene function in early embryos as well as human cell lines. Importantly, insertion of this cistronic construct into replication-incompetent avian retroviruses allowed us to couple gene knockouts with long-term lineage analysis. We demonstrate the application of our newly engineered constructs and viruses by perturbing β-catenin in vitro and Sox10, Pax6 and Pax7 in the neural crest, retina, and neural tube and segmental plate in vivo, respectively. Together, this approach enables genes of interest to be knocked out in identifiable cells in living embryos and can be broadly applied to numerous genes in different embryonic tissues.

Indexed as

CRISPR-Cas SystemsAnimalsChick EmbryoGene EditingGene Expression Regulation, DevelopmentalGene Knockout TechniquesNeural CrestPAX6 Transcription FactorPAX7 Transcription FactorPlasmidsRNA, Guide, CRISPR-Cas SystemsSOXE Transcription FactorsPAX6 Transcription FactorPAX7 Transcription FactorRNA, Guide, CRISPR-Cas SystemsSOXE Transcription FactorsChick embryologyClonal analysisCRISPR-Cas9Genome editingMigrationNeural crestRetroviruses

Identifiers

PMID33688075
PMCPMC8077534
OpenAlexW3134929960

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.