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RNA Interference in Plant Biology: New Tools for an Old Favorite

Posted by Guest Blogger on Oct 27, 2020 9:15:00 AM

This post was contributed by guest blogger, Robert Orr, who recently received a Ph.D. in Biology and Biotechnology from Worcester Polytechnic Institute.

What is RNAi?

The loss-of-function (LOF) experiment functions as the building block of our understanding of complex biological processes. Many tools exist to perturb biological function in a direct or unbiased way at the DNA, RNA, or protein level. The “correct” choice of tool requires careful balancing of the inherent advantages and limitations of any technique in the context of the biological question. For example, while gene knockouts have long been considered the “gold-standard” for LOF studies, the high gene copy number found in plants makes traditional knockouts unattractive from a practical perspective. Therefore, techniques that function downstream of DNA, such as RNA interference, can reversibly exert their effect independent of gene copy number.

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Topics: Plant Biology, Other Plasmid Tools, Plasmids

Save Time with Transient Plant Leaf Transformations

Posted by Guest Blogger on Jul 25, 2019 8:03:52 AM

This post was contributed by Samuel Mortensen, a PhD candidate at Northeastern University.

Working with plants doesn’t always have to be a time-consuming process. While developing transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant can take even longer, a transient plant leaf transformation process could save the plant biologist some time… months, in fact.

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Topics: Plant Biology, Molecular Biology Protocols and Tips, Other

CRISPR-mediated Plant Base Editors

Posted by Guest Blogger on Jan 3, 2019 8:35:29 AM

This post was contributed by Kutubuddin Molla, a Fulbright Visiting Scholar at the Pennsylvania State University.

Imagine you are dealing with a defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can CRISPR be applied to fix the defective base precisely?

Until 2016, precise single base changes were only possible through exploiting the homology-directed repair (HDR) pathway which occurs in cells at low frequencies and relies on the efficient delivery of donor DNA to facilitate repair. Since the development of CRISPR-mediated base editing (BE), these types of repairs can now be done more efficiently than before. A base editor precisely changes a single base with an efficiency typically ranging from 25-75%, while the success of precise change via HDR limited to 0-5%. This blog post covers a brief review of different basic BE technologies and their adaptation for plant genome editing.
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Topics: CRISPR, Plant Biology, Base Editing

Tips for arabidopsis transformation

Posted by Guest Blogger on Oct 25, 2018 9:23:48 AM

This post was contributed by Laura Lee, a graduate student at Stanford University.

Arabidopsis is a fantastic model organism for many reasons, not the least of which is ease of transformation. There are many motivations to generate transgenic Arabidopsis, from studying transcriptional and translational dynamics of genes and proteins in living plants, to complementing mutant phenotypes. Arabidopsis is amenable to the floral drip or dip transformation method. The general steps for this method include:

  • Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a plant pathogenic species that stably integrates transfer DNA (tDNA) into the genomes of the plants it attacks
  • Growing the transformed agrobacterium culture
  • Dipping your plant’s flowers in the agrobacterium culture to allow for tDNA insertions into the plant’s germline
  • Selecting for seeds that have the tDNA insertions (usually via seed growth on antibiotic-containing media)
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Topics: Plant Biology, Molecular Biology Protocols and Tips, Plasmids

Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

Posted by Guest Blogger on Apr 12, 2018 9:02:19 AM

This post was contributed by guest blogger Courtney Price, the Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for Applied Plant Sciences at The Ohio State University.

Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two global stock centers for Arabidopsis thaliana (Arabidopsis). Our mission is to collect, preserve, reproduce and distribute seeds, DNA and other resources for Arabidopsis and related species. Located at The Ohio State University in Columbus, Ohio, ABRC ships more than 100,000 samples to researchers and educators in 60 countries each year.

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Topics: Scientific Sharing, Plant Biology, Material Sharing

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