cDNA (complementary DNA)とは？cDNAは、センス鎖か、アンチセンス鎖か？

cDNAとは、mRNAを鋳型として逆転写酵素により合成したDNA（一重鎖）のことです。cDNAはcomplemantary DNA to mRNAという意味です。通常、実験するうえでcDNAを一重鎖のままにしておくことはなく、cDNAを鋳型としてDNA合成酵素を働かせて二重鎖を得ます。そうしないと、増やしたりできないので。最初につくった一本鎖のcDNAはアンチセンス鎖ですが、それを鋳型にして二重鎖にするために合成した方は、センス鎖になります。cDNAクローニングの論文で示される塩基配列は、センス鎖の塩基配列です。cDNAはmRNAを鋳型にして作った一本鎖DNAという言い方を見ると、論文でcDNA配列といったときの配列とは逆になるのでは？という疑問が湧いて、混乱を招きそうです。そのあたりを、ハッキリさせておきます。

ChatGPTに説明してもらいました。

cDNA is initially single-stranded after it is synthesized from mRNA, and there’s a subtle distinction when considering single- and double-stranded DNA.

Here’s a more precise breakdown of what happens:

  1. Single-stranded cDNA:
    • cDNA is synthesized from the mRNA template by reverse transcriptase, making it complementary to the mRNA. Since mRNA is complementary to the anti-sense (template) strand of genomic DNA, the single-stranded cDNA is complementary to the mRNA, which essentially makes it analogous to the anti-sense strand of genomic DNA.
  2. Double-stranded cDNA:
    • If the cDNA is made into a double-stranded molecule (which is often the case in practical applications), then the second strand is synthesized to be complementary to the first (single-stranded) cDNA. This newly synthesized strand is now complementary to the cDNA and, as a result, matches the sequence of the original mRNA (or the sense strand of genomic DNA).

Thus, when cDNA is single-stranded, it is effectively the anti-sense strand (complementary to mRNA). However, once you make it double-stranded, the complementary strand matches the sense strand of the original genomic DNA.

Summary:

  • Single-stranded cDNA corresponds to the anti-sense strand of the genomic DNA.
  • When cDNA becomes double-stranded, the newly synthesized second strand is the sense strand, which matches the original genomic DNA’s sense strand (with T instead of U).

通常、cDNA合成というときは、 first-strand synthesis, second-strand synthesisといった言い方をします。そのあたりをChatGPTに説明してもらいました。

In the context of cDNA synthesis:

  • The strand from the “first-strand synthesis” of cDNA is the one synthesized directly from the mRNA template. Since mRNA is complementary to the anti-sense strand of the original genomic DNA, this first-strand cDNA is complementary to the mRNA and is therefore analogous to the anti-sense strand of the original DNA.
  • The strand from the “second-strand synthesis” of cDNA is synthesized using the first-strand cDNA as a template. This second strand will be complementary to the first strand, which means it will have the same sequence as the original sense strand of the genomic DNA (except for thymine (T) replacing uracil (U) found in mRNA).

In summary:

  • The first-strand cDNA corresponds to the anti-sense strand.
  • The second-strand cDNA corresponds to the sense strand.