RNA interference-mediated gene knockdown within specific cell types

Changqing Zhang, David W Galbraith

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

In plants, RNA interference (RNAi)-induced gene silencing can spread from the initiation site to nearby cells. The silencing signal moves from cell-to-cell through plasmodesmata and, over long distances, through the phloem. In this study, we employed a nuclear-localized GFP fusion protein to visualize the pattern of gene silencing induced by three different transgenes expressing double-stranded RNA (dsRNA) in Arabidopsis root tips. In all cases, we found that dsRNA-induced silencing did not spread from the silencing initiation site to adjacent cells. In the first set of experiments, in a transgenic background expressing nuclear-localized GFP within a contiguous cell layer that included endodermis, cortex/endodermis (joint) initial (CEI) cells and the quiescent center (QC) cells, expression of the marker gene was silenced specifically in the QC cells without affecting gene expression in the adjacent CEI and endodermal cells. The next two sets of experiments examined the knockdown of two endogenous genes. We observed that silencing was completely restricted to the QC and endodermal cells within which the dsRNA transgenes were expressed. Overall, these results accentuate one important aspect of RNAi-induced gene silencing, that it can be cell autonomous, and demonstrated the feasibility of selective gene knockdown within specific cell types.

Original languageEnglish (US)
Pages (from-to)169-176
Number of pages8
JournalPlant Molecular Biology
Volume80
Issue number2
DOIs
StatePublished - Sep 2012

Fingerprint

Gene Knockdown Techniques
RNA Interference
RNA interference
genes
cells
Double-Stranded RNA
double-stranded RNA
Gene Silencing
gene silencing
endodermis
Transgenes
transgenes
Plasmodesmata
Plant RNA
Phloem
Gene Expression
plasmodesmata
Meristem
Arabidopsis
root tips

Keywords

  • GFP
  • RNAi
  • Silencing signal

ASJC Scopus subject areas

  • Plant Science
  • Agronomy and Crop Science
  • Genetics

Cite this

RNA interference-mediated gene knockdown within specific cell types. / Zhang, Changqing; Galbraith, David W.

In: Plant Molecular Biology, Vol. 80, No. 2, 09.2012, p. 169-176.

Research output: Contribution to journalArticle

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