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使用CRISPR/Cas9对种子繁殖和营养繁殖中的基因编辑结果进行全基因组评估。

Genome-wide evaluation of gene editing outcomes using CRISPR/Cas9 in seed propagated and vegetatively propagated .

作者信息

Jayakody Thilani B, Zarka Daniel, Cho Keun Ho, Jensen Jacob, Sikora Samantha, Buell C Robin, Douches David S, Nadakuduti Satya Swathi

机构信息

Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, United States.

Environmental Horticulture Department, University of Florida, Gainesville, FL, United States.

出版信息

Front Plant Sci. 2024 Nov 26;15:1496861. doi: 10.3389/fpls.2024.1496861. eCollection 2024.

DOI:10.3389/fpls.2024.1496861
PMID:39659410
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11628256/
Abstract

CRISPR/Cas9 is the most popular genome editing platform for investigating gene function or improving traits in plants. The specificity of gene editing has yet to be evaluated at a genome-wide scale in seed-propagated (L.) Crantz (camelina) or clonally propagated L. (potato). In this study, seven potato and nine camelina stable transgenic Cas9-edited plants were evaluated for on and off-target editing outcomes using 55x and 60x coverage whole genome shotgun sequencing data, respectively. For both potato and camelina, a prevalence of mosaic somatic edits from constitutive Cas9 expression was discovered as well as evidence of transgenerational editing in camelina. CRISPR/Cas9 editing provided negligible off-target activity compared to background variation in both species. The results from this study guide deployment and risk assessment of genome editing in commercially relevant traits in food crops.

摘要

CRISPR/Cas9是用于研究植物基因功能或改良植物性状的最流行的基因组编辑平台。在种子繁殖的荠蓝（Camelina sativa (L.) Crantz）或克隆繁殖的马铃薯（Solanum tuberosum L.）中，尚未在全基因组范围内评估基因编辑的特异性。在本研究中，分别使用覆盖度为55x和60x的全基因组鸟枪法测序数据，对7株马铃薯和9株荠蓝稳定转基因Cas9编辑植株的靶向和脱靶编辑结果进行了评估。对于马铃薯和荠蓝，均发现了组成型Cas9表达导致的嵌合体细胞编辑普遍存在，以及荠蓝中跨代编辑的证据。与两个物种的背景变异相比，CRISPR/Cas9编辑产生的脱靶活性可忽略不计。本研究结果为粮食作物商业相关性状的基因组编辑的应用和风险评估提供了指导。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/08e1d25ad6d0/fpls-15-1496861-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/ca981edd155b/fpls-15-1496861-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/94db92701a13/fpls-15-1496861-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/42cf877244d6/fpls-15-1496861-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/cc33e42b8225/fpls-15-1496861-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/b0b50a688c94/fpls-15-1496861-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/08e1d25ad6d0/fpls-15-1496861-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/ca981edd155b/fpls-15-1496861-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/94db92701a13/fpls-15-1496861-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/42cf877244d6/fpls-15-1496861-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/cc33e42b8225/fpls-15-1496861-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/b0b50a688c94/fpls-15-1496861-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fe3/11628256/08e1d25ad6d0/fpls-15-1496861-g006.jpg

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