《Table 1–Average amylose contents of selected T1homozygous lines.》

  提示：宽带有限、当前游客访问压缩模式

本系列图表出处文件名：随高清版一同展现

《Systematic identification of endogenous RNA polymerase Ⅲ promoters for efficient RNA guidebased genome editing technologies in maize》

1. 获取 高清版本忘记账户？点击这里登录

1. 下载图表忘记账户？点击这里登录

aPairwise t-test of generated lines and corresponding wild types.

The in vitro data suggested that all seven Pol III promoters operated on the CRISPR/Cas9 construct，but with mutation varying from 3.4%（U6-1）to over 21.0%（U6-6），indicating different activities.In Fig.4 a detailed example of RNA-guided Cas9 targeting the Zm Wx1 locus is shown，wherein a singleguide RNA was constructed that was driven by the promoter U6-2.An expression cassette of RNA-guided Cas9（Fig.4A）was designed to target the Zm Wx1 locus（Fig.4B）.Among the 66 T0transformation-positive events from three rounds of transformation，the targeted mutation efficiencies ranged from48.5%to 97.1%.Among them，36 insertions of a single T and 15of a single A comprised the majority of the mutation types.The mutation frequencies were high，such that 32 of 66recessive wx1 mutant homozygotes exhibited a waxy phenotype，in which amylose could not be detected in the kernels of T0 plants.Selfing those families revealed that the targeted mutations all heritably produced recessive wx1 homozygous phenotypes.The kernel endosperm starch and pollen phenotypes of four randomly selected T1 families（CNW-D19，CNW-D5，CNW-D21，and CNW-D45）were scored based on KIstaining（Fig.5）and amylose content determination（Table1）.In addition to the visible phenotypes of the KI staining of both the kernel endosperm starch particles and pollen cells，the amylose contents of the mutants were low to undetectable，compared with amylose contents of approximately 18%in the wild type（Table 1）.These in vivo data for Zm Wx1 suggest that the U6-2 promoter could be used as a key element of CRISPR/Cas genome editing tools and will exhibit robust activity.