Is it possible to use a biologically active Telemorease Elongation Reverse Transcriptase (TERT) in the place of the Reverse Transcriptase (RT) for quantitative Reverse Transcriptase pcr (qRT-pcr)? Yes I do know that it would seemingly have no advantages. What kind of reverse transcriptase is used in qRT-pcr is there a standard like taq dna pol is used for qpcr?
2 Answers
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hTERT is incapable of replacing common reverse transcriptases like MuLV or SuperScript.
hTERT is specialized to bind with its own RNA (hTR aka TERC), and not any given RNA. hTERT uses hTR as a template to extend a DNA strand with the same repeats (TTAGGG). (Cairney & Keith 2008). For qPCR it's needed to transcribe RNA into the same sequence of DNA, not something repetitive.
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$\begingroup$ Could you provide links describing the biochemistry of MuLV and SuperScript? On the face of it I only get the gist that they are for reverse transcription. $\endgroup$– GalenSep 10, 2022 at 17:28
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1$\begingroup$ Zhao et al. 2018 is one free article (so many other articles are behind a paywall). It‘s interesting since it discusses an RT that‘s not isolated from a virus (like every coomercial RT out there), but actually from a transposon (which are like ancient viral elements in the genome of every animal). It also compares it to SuperscriptIV, which is the pinnacle of enzyme beauty. Have fun reading. $\endgroup$– markurSep 10, 2022 at 23:23
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What kind of reverse transcriptase is used in qRT-pcr is there a standard like taq dna pol is used for qPCR
M-MuLV (Monoley murine leukemia virus) reverse transcriptase
Some RT enzymes are engineered to remove the RNAseH domain so that template RNA persists for more rounds of RT.
Is it possible to use a biologically active Telemorease Elongation Reverse Transcriptase (TERT) in the place of the Reverse Transcriptase
Perhaps it can catalyse an RT reaction in-vitro. But commercial enzymes need to be efficient and I think using TERT won't be that advantageous.
