Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells

Abstract

Morpholino oligonucleotides (MO) are an innovative tool that provides a means for examining and modifying gene expression outcomes by antisense interaction with targeted RNA transcripts. The site-specific nature of their binding facilitates focused modulation to alter splice variant expression patterns. Here we describe the steric-blocking of human telomerase reverse transcriptase (hTERT) Δα and Δβ splice variants using MO to examine cellular outcomes related to pluripotency and differentiation in human embryonic stem cells.

The role of telomerase reverse transcriptase in the mitochondrial protective functions of Angiotensin-(1–7) in diabetic CD34+ cells

“…Selective steric blockade of β-TERT splicing was accomplished by using oligonucleotide sequence (5′CACCTGGCCACCTGAC TCACTTGCC3′) that is complimentary to specific exon/intron boundaries of the pre-mRNA sequence of TERT (e8/i8) (5′GGCAAgtgag tcaggtggccaggtg3′), and oligos were delivered in form of PMOs (Gene Tools, Inc.). [ 25 , 61 ] As observed above, FL-TERT expression was lower in DB-CD34 + cells compared with ND cells ( P < 0.01, n = 5) ( Fig. 6A ) which was increased by β-TERT-PMO ( P < 0.05, n = 5) ( Fig.…”