Template switching within exons 3 and 4 of KV11.1 (HERG) gives rise to a 5′ truncated cDNA

S. Ro, S. H. Kang, A. M. Farrelly, T. Ordog, R. Partain, Neal Fleming, K. M. Sanders, J. L. Kenyon, K. D. Keef

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

KV11.1 (HERG) channels contribute to membrane potential in a number of excitable cell types. We cloned a variant of KV11.1 from human jejunum containing a 171 bp deletion spanning exons 3 and 4. Expression of a full-length cDNA clone containing this deletion gave rise to protein that trafficked to the cell membrane and generated robust currents. The deletion occurred in a G/C-rich region and identical sequence elements of UGGUGG were located at the deletion boundaries. In recent studies these features have been implicated to cause deletions via template switching during cDNA synthesis. To examine this possibility we compared cDNAs from human brain, heart, and jejunum synthesized at lower (42 °C) and higher temperatures (70 °C). The 171 bp deletion was absent at the higher temperature. Our results suggest that the sequence and secondary structure of mRNA in the G/C rich region leads to template switching producing a cDNA product with a 171 bp deletion.

Original languageEnglish (US)
Pages (from-to)1342-1349
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume345
Issue number4
DOIs
StatePublished - Jul 14 2006

Fingerprint

Exons
Complementary DNA
Jejunum
Temperature
Cell membranes
Membrane Potentials
Brain
Clone Cells
Cell Count
Cell Membrane
Membranes
Messenger RNA
Proteins

Keywords

  • Brain
  • Cardiac
  • Gastrointestinal
  • HERG channel
  • RT slippage
  • Template switching

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Template switching within exons 3 and 4 of KV11.1 (HERG) gives rise to a 5′ truncated cDNA. / Ro, S.; Kang, S. H.; Farrelly, A. M.; Ordog, T.; Partain, R.; Fleming, Neal; Sanders, K. M.; Kenyon, J. L.; Keef, K. D.

In: Biochemical and Biophysical Research Communications, Vol. 345, No. 4, 14.07.2006, p. 1342-1349.

Research output: Contribution to journalArticle

Ro, S, Kang, SH, Farrelly, AM, Ordog, T, Partain, R, Fleming, N, Sanders, KM, Kenyon, JL & Keef, KD 2006, 'Template switching within exons 3 and 4 of KV11.1 (HERG) gives rise to a 5′ truncated cDNA', Biochemical and Biophysical Research Communications, vol. 345, no. 4, pp. 1342-1349. https://doi.org/10.1016/j.bbrc.2006.05.032
Ro, S. ; Kang, S. H. ; Farrelly, A. M. ; Ordog, T. ; Partain, R. ; Fleming, Neal ; Sanders, K. M. ; Kenyon, J. L. ; Keef, K. D. / Template switching within exons 3 and 4 of KV11.1 (HERG) gives rise to a 5′ truncated cDNA. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 345, No. 4. pp. 1342-1349.
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