Data Mining by Pluralistic Approach on CRISPR Gene Editing in Plants.

Tanushri Kaul,Sonia Khan Sony,Murugesh Eswaran,Rachana Verma,Rashmi Kaul,Pranjal Yadava,Pawan Kumar Agrawal,Krishnamurthy M Sathelly,Arulprakash Thangaraj,Nitya Meenakshi Raman

doi:10.3389/fpls.2019.00801

Abstract

Genome engineering by site-specific nucleases enables reverse genetics and targeted editing of genomes in an efficacious manner. Contemporary revolutionized progress in targeted-genome engineering technologies based on Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-related RNA-guided endonucleases facilitate coherent interrogation of crop genome function. Evolved as an innate component of the adaptive immune response in bacterial and archaeal systems, CRISPR/Cas system is now identified as a versatile molecular tool that ensures specific and targeted genome modification in plants. Applications of this genome redaction tool-kit include somatic genome editing, rectification of genetic disorders or gene therapy, treatment of infectious diseases, generation of animal models, and crop improvement. We review the utilization of these synthetic nucleases as precision, targeted-genome editing platforms with the inherent potential to accentuate basic science “strengths and shortcomings” of gene function, complement plant breeding techniques for crop improvement, and charter a knowledge base for effective use of editing technology for ever-increasing agricultural demands. Furthermore, the emerging importance of Cpf1, Cas9 nickase, C2c2, as well as other innovative candidates that may prove more effective in driving novel applications in crops are also discussed. The mined data has been prepared as a library and opened for public use at www.lipre.org.

Highlights

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system is largely involved in conferring resistance to genetic transformants, and viruses, if present in surrounding environment, rendering a type of acquired immunity
The CRISPR system has been estimated in a record of 87 plants until 2017 revealing that the adaptation of this genome editing technology has been achieved with minimum off-target mutations by many research groups worldwide
Among the three loci targeted in the maize genome by using dmc1 gene promoter with the U3 promoter for the sgRNA and Cas9, high efficiency of gene editing was observed in T0 plants with 66% stable transmission of the mutation to the T1 generation

Summary

Introduction

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system is largely involved in conferring resistance to genetic transformants, and viruses, if present in surrounding environment, rendering a type of acquired immunity. The main strategy employed to increase the editing efficiency of the CRISPR/Cas9 system is the use of strong promoters and to avoid usage of a low-scored guide sequence (Hu et al, 2018). The CRISPR system has been estimated in a record of 87 plants until 2017 revealing that the adaptation of this genome editing technology has been achieved with minimum off-target mutations by many research groups worldwide.

Results

Conclusion