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Frequently asked questions (FAQs)

Is the PAM sequence part of the CRISPR-Cas9 crRNA construct?

Sep 19, 2017, 18:32 PM

No. The PAM sequence is located on the non-complementary strand. That is, it is on the strand of DNA that contains the same DNA sequence as the target crRNA [1]. The PAM sequence should not be included in the design of the crRNA.

The most commonly used Cas9 nuclease, derived from S. pyogenes, recognizes a PAM sequence of NGG that is found directly downstream of the target sequence in the genomic DNA, on the non-target strand.

Want to Learn More?

Download The CRISPR Basics Handbook that covers applications of CRISPR technology from guide RNA design to data analysis.

The latest CRISPR abbreviations and definitions in a single list!

Design custom solutions for CRISPR genome editing.

Reference

  1. Anders C, Niewoehner O, et al. (2014) Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature. 513(7519):569–573.
Tags :
  • alt-r
  • cas9
  • crispr
  • PAM
Categories :
  • Alt-R CRISPR nucleases
  • Alt-R CRISPR-Cas9
  • CRISPR genome editing
  • FAQs
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