Evaluation of a novel short polyadenylation signal as an alternative to the SV40 polyadenylation signal.
The soluble neuropilin-1 (sNRP-1) gene employs an extremely short dual function polyadenylation (pA) signal/stop codon that is efficient for termination of gene transcription and translation in vivo. However, the functionality and usefulness of this signal in regard to other genes is unknown. This quantitative study compares the levels of humanized Renilla green fluorescent protein (hrGFP) mRNA polyadenylated with either the sNRP-1 pA signal or the much larger, more widely used SV40 pA signal. We show that the overall starting copy number of hrGFP for equally loaded RNAs is equivalent between the two groups. Our data show little to no difference between levels of mRNA generated by the sNRP-1 polyadenylation signal and the SV40 polyadenylation signal despite a remarkable size difference in signal length. An extremely short polyadenylation signal could potentially alleviate gene insert size restrictions associated with cloning or with therapeutic vectors such as adeno-associated virus.
Casey Eye Institute, Oregon Health and Science University, 3375 SW Terwilliger Blvd, Portland, 97239-4197, USA.
Trevor J
TJ
McFarland
Yi
Y
Zhang
La-ongsri
LO
Atchaneeyaskul
Peter
P
Francis
J Timothy
JT
Stout
Binoy
B
Appukuttan
D001483:Base Sequence; D003001:Cloning, Molecular; D018388:Codon, Terminator; D000229:Dependovirus / Q000235:genetics; D005091:Exons; D016678:Genome; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D039942:Neuropilin-1 / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics*; D026723:Polyadenylation; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000378:metabolism
D018388:Codon, Terminator; D009842:Oligopeptides; D020033:Protein Isoforms; D012333:RNA, Messenger; C402304:SV40 nuclear localization peptide; D039942:Neuropilin-1; D049452:Green Fluorescent Proteins