Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Jan 28;48(6):3228–3243. doi: 10.1093/nar/gkz1240

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

PMC Copyright notice

Figure 3. — Cleavage efficiency for spacers with different upstream and downstream sequences. (A) According to our hypothesis, the Sp1, Sp4 and Sp12 will have efficient processing for both repeats. The Sp8 causes misfolding of the pseudoknot and thereby impedes the processing of the first repeat. (B) In vitro pre-crRNA processing of Sp1, Sp4, Sp8 and Sp12 at different time intervals. The marker lane on the left shows three bands from the low range ssRNA ladder (NEB). The bands of 127 nt, 66 nt and 37 nt indicate processing of the pre-crRNA that would lead to a targeting RNP. The processing of the second repeat (103 nt, 61 nt) gives rise to a non-targeting RNP. (C) Two relatively inefficient spacers were tested in different pre-crRNA architectures (pTarget1 - pTarget6): Sp8 (red bars) and Sp12-variant [cUA] (blue bars). Average values from three biological replicates are shown, with error bars representing SD. A 20 nt leader-end is shown in orange, a full or mature repeat in blue, a 24 nt spacer sequence in red, a SalI restriction site in yellow, a spacing sequence in green and the terminator is shown in purple. Spacers were flanked on the 5′ end with a leader-end sequence and full repeat, or they were flanked by a mature repeat only. Within each category were spacers containing various downstream sequences. Sequences such as, only a terminator (|T|), a mature repeat-spacing sequence-terminator (MR) and a full repeat-spacing sequence-terminator (FR).