<div style="text-align:justify">Background: In eukaryotes, Histone acetylation and deacetylation plays a prominent role in controlling gene expression and chromatin structure modifications. Both Histone acetyltransferases (HAT) and Histone deacetylases (HDACs) work in opposition to regulate chromatin acetylation. Reduced levels of histone H4 acetylation and increased susceptibility to the rice disease M. oryzae result from overexpression of HDT701 in rice. By changing the levels of histone H4 acetylation in defense-related and pattern recognition receptor (PRR) genes in rice, HDT701 reduces the activity of innate immunity which promotes resistance to M. oryzae. Crispr cas9 technology was created to change genes and modify characteristics, as well as to produce resistance to a variety of infections by focusing on possible biomolecules involved in plant defense mechanisms. Therefore the present study was aimed to design single guided RNA (sgRNA) and predict the gene mutational position in the HDT gene in RPBio-226 rice cv. 
 
Methods: To begin with, the DNA was isolated using CTAB method. Specific pair of primers was designed from the reference gene for amplification of HDT gene. In addition to, the PCR product was sequenced and the resulting sequence was applied to the creation of sgRNA. Furthermore, CHOPCHOP is a Bioinformatic search tool used to identify CRISPR&ndash;Cas single guide RNA (sgRNA) targets. 
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<div>Result: The isolated genomic DNA was quantified using nanodrop and found that the concentration of the DNA was 800-1000 ng/ul with the purity of 1.8. The full gene was amplified with OsHDT701 gene primers and sequenced. Based on the OsHDT701 gene sequence, oligo single guide RNA (sgRNA) was generated by using the http://chopchop.cbu.uib programme. The target site for designing sgRNA was found from 168 basepair to 190 basepair with the deletion of a nucleotide at 174th position.</div>

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In-silico Analysis for Prediction of Mutational Position and Designing of sgRNA for HDT701 Gene in Indian Rice cv. RPBio-226

Background: Genome editing is a group of technologies that has the ability to change an organism&rsquo;s DNA. These technologies allow genetic material to be added, removed, or altered at particular locations in the genome. Several approaches to genome editing have been developed. A well-known one is called CRISPR/Cas9, which is short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9. Editing through CRISPR focuses mainly on the Protospacer-adjacent motif (PAM), a crucial region to identify the sgRNAs to target the desired gene or region of DNA. So designing precise single-guide RNAs (sgRNAs) to minimize off-target effects is critical for the success of gene editing without undesired results. 
 
Methods: Hence, in this article, three different online tools are used to design sgRNA to target the prolactin (PRL) gene to knock out. The Gallus gallus Prolactin (PRL) gene sequence is retrieved from NCBI and used for further downstream application. However, all three web tools may vary in design specifications and parameter choices, visualization, downstream analysis functionality, etc.&nbsp; 
 
Result: While keeping a straightforward and interactive interface and running with default parameters, all web tools also accept a variety of advanced choices for more specialized searches. This maximizes user flexibility. Three tools produced several sgRNAs that satisfied various criteria for precise gene editing to boost the efficacy of the target prolactin gene. These online tools use a robust approach to identify off-target locations and the results are displayed in an interactive table and within the gene architecture.

CRISPR/Cas Genome Editing Single Guide RNA (sgRNA) Design using Three Different Web Tool Platforms

<div>Clustered regularly interspaced short palindromic repeats/CRISPR associated nuclease 9 (CRISPR-Cas9) system is a rapid technology for gene editing. CRISPR-Cas9 is an RNA guided gene editing tool where Cas9 acts as endonuclease by cutting the target DNA strand. Double Stranded Breaks (DBS) can be repaired by non-homologous end joining (NHEJ) and homology-directed repair (HDR). The NHEJ employs DNA ligase IV to rejoin the broken ends which cause insertion or deletion mutations, whereas HDR repairs the DSBs based on a homologous complementary template and results in perfect repair of broken ends. CRISPR-Cas9 impart diverse advantageous features in contrast with the conventional methods. In this review article, we have discussed CRISPR-Cas9 based genome editing along with its mechanism of action and role in crop improvement.</div>

In-silico Analysis for Prediction of Mutational Position and Designing of sgRNA for HDT701 Gene in Indian Rice cv. RPBio-226

CRISPR/Cas Genome Editing Single Guide RNA (sgRNA) Design using Three Different Web Tool Platforms

CRISPR-Cas9: A Genome Editing Tool in Crop Plants: A Review