Simple monitoring of endocrine-disrupting chemicals using transgenic Arabidopsis plants expressing medaka estrogen receptor

Stoykova, Petya; Ohkawa, Hideo; Inui, Hideyuki

doi:10.1016/j.chemosphere.2021.131633

Cited by 4 publications

(1 citation statement)

References 45 publications

(61 reference statements)

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“…Fluorescent proteins provide an interesting option in this regard. The green fluorescent protein (GFP) is a prevalent reporter gene used for the visual detection of transgenic tissues (e.g., [42,43]). However, it is not suitable for lignified or flavone-containing tissues as their autofluorescence emission overlaps with GFP, and this background interference makes it difficult for the in vivo detection of GFP.…”

Section: Hairy Root Transformationmentioning

confidence: 99%

CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean

Niazian

Belzile

Torkamaneh

2022

Plants

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Sequence and expression data obtained by next-generation sequencing (NGS)-based forward genetics methods often allow the identification of candidate causal genes. To provide true experimental evidence of a gene’s function, reverse genetics techniques are highly valuable. Site-directed mutagenesis through transfer DNA (T-DNA) delivery is an efficient reverse screen method in plant functional analysis. Precise modification of targeted crop genome sequences is possible through the stable and/or transient delivery of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (CRISPR/Cas) reagents. Currently, CRISPR/Cas9 is the most powerful reverse genetics approach for fast and precise functional analysis of candidate genes/mutations of interest. Rapid and large-scale analyses of CRISPR/Cas-induced mutagenesis is achievable through Agrobacterium rhizogenes-mediated hairy root transformation. The combination of A. rhizogenes hairy root-CRISPR/Cas provides an extraordinary platform for rapid, precise, easy, and cost-effective “in root” functional analysis of genes of interest in legume plants, including soybean. Both hairy root transformation and CRISPR/Cas9 techniques have their own complexities and considerations. Here, we discuss recent advancements in soybean hairy root transformation and CRISPR/Cas9 techniques. We highlight the critical factors required to enhance mutation induction and hairy root transformation, including the new generation of reporter genes, methods of Agrobacterium infection, accurate gRNA design strategies, Cas9 variants, gene regulatory elements of gRNAs and Cas9 nuclease cassettes and their configuration in the final binary vector to study genes involved in root-related traits in soybean.

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Section: Hairy Root Transformationmentioning

confidence: 99%