PMID- 23894227
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20211021
IS  - 1424-8247 (Print)
IS  - 1424-8247 (Electronic)
IS  - 1424-8247 (Linking)
VI  - 6
IP  - 3
DP  - 2013 Mar 14
TI  - Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers.
PG  - 295-319
AB  - Recent clinical trials of small interfering RNAs (siRNAs) highlight the need for 
      robust delivery technologies that will facilitate the successful application of 
      these therapeutics to humans. Arguably, cell targeting by conjugation to 
      cell-specific ligands provides a viable solution to this problem. Synthetic RNA 
      ligands (aptamers) represent an emerging class of pharmaceuticals with great 
      potential for targeted therapeutic applications. For targeted delivery of siRNAs 
      with aptamers, the aptamer-siRNA conjugate must be taken up by cells and reach 
      the cytoplasm. To this end, we have developed cell-based selection approaches to 
      isolate aptamers that internalize upon binding to their cognate receptor on the 
      cell surface. Here we describe methods to monitor for cellular uptake of 
      aptamers. These include: (1) antibody amplification microscopy, (2) 
      microplate-based fluorescence assay, (3) a quantitative and ultrasensitive 
      internalization method ("QUSIM") and (4) a way to monitor for cytoplasmic 
      delivery using the ribosome inactivating protein-based (RNA-RIP) assay. 
      Collectively, these methods provide a toolset that can expedite the development 
      of aptamer ligands to target and deliver therapeutic siRNAs in vivo.
FAU - Hernandez, Luiza I
AU  - Hernandez LI
AD  - Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA.
FAU - Flenker, Katie S
AU  - Flenker KS
FAU - Hernandez, Frank J
AU  - Hernandez FJ
FAU - Klingelhutz, Aloysius J
AU  - Klingelhutz AJ
FAU - McNamara, James O 2nd
AU  - McNamara JO 2nd
FAU - Giangrande, Paloma H
AU  - Giangrande PH
LA  - eng
GR  - R01 CA138503/CA/NCI NIH HHS/United States
GR  - R21 DE019953/DE/NIDCR NIH HHS/United States
PT  - Journal Article
PL  - Switzerland
TA  - Pharmaceuticals (Basel)
JT  - Pharmaceuticals (Basel, Switzerland)
JID - 101238453
PMC - PMC3722562
MID - NIHMS480193
OTO - NOTNLM
OT  - RNA aptamers
OT  - cell-SELEX
OT  - cell-internalizing aptamers
OT  - siRNA delivery
OT  - targeted delivery
EDAT- 2013/07/31 06:00
MHDA- 2013/07/31 06:01
PMCR- 2013/03/01
CRDT- 2013/07/30 06:00
PHST- 2013/07/30 06:00 [entrez]
PHST- 2013/07/31 06:00 [pubmed]
PHST- 2013/07/31 06:01 [medline]
PHST- 2013/03/01 00:00 [pmc-release]
AID - pharmaceuticals-06-00295 [pii]
AID - 10.3390/ph6030295 [doi]
PST - ppublish
SO  - Pharmaceuticals (Basel). 2013 Mar 14;6(3):295-319. doi: 10.3390/ph6030295.