Enhancing Endosomal Escape and Gene Regulation Activity for Spherical Nucleic Acids.

ABSTRACT: The therapeutic potential of small interfering RNAs (siRNAs) is limited by their poor stability and low cellular uptake. When formulated as spherical nucleic acids (SNAs), siRNAs are resistant to nuclease degradation and enter cells without transfection agents with enhanced activity compared to their linear counterparts; however, the gene silencing activity of SNAs is limited by endosomal entrapment, a problem that impacts many siRNA-based nanoparticle constructs. To increase cytosolic delivery, SNAs are formulated using calcium chloride (CaCl₂ ) instead of the conventionally used sodium chloride (NaCl). The divalent calcium (Ca²⁺ ) ions remain associated with the multivalent SNA and have a higher affinity for SNAs compared to their linear counterparts. Importantly, confocal microscopy studies show a 22% decrease in the accumulation of CaCl₂ -salted SNAs within the late endosomes compared to NaCl-salted SNAs, indicating increased cytosolic delivery. Consistent with this finding, CaCl₂ -salted SNAs comprised of siRNA and antisense DNA all exhibit enhanced gene silencing activity (up to 20-fold), compared to NaCl-salted SNAs regardless of sequence or cell line (U87-MG and SK-OV-3) studied. Moreover, CaCl₂ -salted SNA-based forced intercalation probes show improved cytosolic mRNA detection.

SUBMITTER: Park J 

PROVIDER: S-EPMC10947971 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature