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siRNA-Mediated RNA Interference in Precision-Cut Tissue Slices Prepared from Mouse Lung and Kidney
Ist Teil von
The AAPS journal, 2017-11, Vol.19 (6), p.1855-1863
Ort / Verlag
New York: Springer US
Erscheinungsjahr
2017
Quelle
MEDLINE
Beschreibungen/Notizen
Small interfering RNA (siRNA)-mediated RNAi interference (RNAi) is a powerful post-transcriptional gene silencing mechanism which can be used to study the function of genes
in vitro
(cell cultures) and
in vivo
(animal models). However, there is a translational gap between these models. Hence, there is a need for novel experimental models that combine the advantages of
in vitro
and
in vivo
models (e.g., simplicity, flexibility, throughput, and representability) to study the effects of siRNA. This need may be addressed by precision-cut tissue slices (PCTS), which represent an
ex vivo
model that mimics the structural and functional characteristics of a whole organ. The goal of this study was to investigate whether self-deliverable siRNA (Accell siRNA) can be used in precision-cut lung slices (PCLuS) and precision-cut kidney slices (PCKS) to achieve RNAi
ex vivo
. PCLuS and PCKS were prepared from mouse tissue, and they were subsequently incubated up to 48 h with no siRNA (untransfected), non-targeting Accell siRNA, or
Gapdh
-targeting Accell siRNA. Significant
Gapdh
mRNA silencing was achieved (PCLuS ~ 55%; PCKS ~ 40%) without compromising the viability and morphology of slices. Fluorescence microscopy confirmed that Accell siRNA diffused into PCLuS and PCKS. Spontaneous inflammation upon incubation was observed in PCLuS and PCKS as shown by a higher mRNA expression of pro-inflammatory cytokines
Il1b
,
Il6
, and
Tnfa
, although Accell siRNA appeared to diminish this response in PCLuS after 24 h. In conclusion, this
ex vivo
transfection model can be used to evaluate the effects of siRNA in relevant biological environments.