Abstract
Global population is predicted to approach 10 billion by 2050, an increase of over 2 billion from today. To meet the demands of growing, geographically and socio-economically diversified nations, we need to diversity and expand agricultural production. This expansion of agricultural productivity will need to occur under increasing biotic, and environmental constraints driven by climate change. Clustered regularly interspaced short palindromic repeats-site directed nucleases (CRISPR-SDN) and similar genome editing technologies will likely be key enablers to meet future agricultural needs. While the application of CRISPR-Cas9 mediated genome editing has led the way, the use of CRISPR-Cas12a is also increasing significantly for genome engineering of plants. The popularity of the CRISPR-Cas12a, the type V (class-II) system, is gaining momentum because of its versatility and simplified features. These include the use of a small guide RNA devoid of trans-activating crispr RNA, targeting of T-rich regions of the genome where Cas9 is not suitable for use, RNA processing capability facilitating simpler multiplexing, and its ability to generate double strand breaks (DSB) with staggered ends. Many monocot and dicot species have been successfully edited using this Cas12a system and further research is ongoing to improve its efficiency in plants, including improving the temperature stability of the Cas12a enzyme, identifying new variants of Cas12a or synthetically producing Cas12a with flexible PAM sequences. In this review we provide a comparative survey of CRISPR-Cas12a and Cas9, and provide a perspective on applications of CRISPR-Cas12 in agriculture.
Highlights
Innovation has always been the driver of agricultural advancement from the earliest days of domestication to today’s machine learning-based genomic selection technologies
The expression of F. novicida and L. bacterium ND2006 nucleases resulted in a high frequency of homologydirected repair (HDR) in rice suggesting a primary advantage of the Cas12a system over Cas9 for targeted gene integration (Begemann et al, 2017)
We have reviewed some of the fundamental differences between the two most widely utilized systems for plant genome engineering and suggest that both Cas9 and Cas12a have unique advantages and disadvantages for genome engineering
Summary
Innovation has always been the driver of agricultural advancement from the earliest days of domestication to today’s machine learning-based genomic selection technologies. Cas12a displays a temperature sensitivity that has limited its utility in plant genome editing (Malzahn et al, 2019; Safari et al, 2019; Swarts, 2019), engineered variants have recently been generated with enhanced activities (Schindele and Puchta, 2020).
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