J. SURESH, R. NAGANNA, B.S.K.NIKHIL, S.SRINIVASA REDDY, AND C.NARENDRA REDDY
Abstract
The recent development of gene-editing technologies, such as Clustered Regularly Interspaced Short Palindromic Repeats and associated protein (CRISPR/Cas), opened new avenues for the development of novel pest control measures. The CRISPR/Cas9 system acts via a ribonucleoprotein complex, where the target recognition lobe of Cas9 directs specific binding to target DNA through interacting with homologous sgRNA and the excision lobe cuts the DNA. CRISPR-containing organisms acquire DNA fragments from invading bacteriophages and plasmids before transcribing them into CRISPR RNAs (crRNAs) to guide cleavage of invading RNA or DNA. The efficiency of target editing is dependent on intrinsic factors specific to each species, the target gene sequence, and the delivery methods of CRISPR gRNA and the Cas nuclease. The CRISPR/Cas9 cargo is delivered in different formats for genome editing in cells; Cas9 may be delivered as a DNA or mRNA molecule encoding for the cas9 gene, or it may be delivered as a functional ribonucleoprotein (RNP) into insect body for the control. In cultures, CRISPR cargo is commonly delivered by physical or non-viral and viral delivery approaches viz., physical approaches; lipofection, electroporation, nucleofection, microinjection and viral vectors such as lentiviruses, adenovirus, and adeno-associated virus (AAV) are broadly used as delivery vehicles of CRISPR cargo for efficient genome editing.