CRISPR technology has been utilized in the agriculture and food sectors for creating probiotic cultures and safeguarding industrial cultures, such as those used in yogurt production, against infections. This technology is also employed to improve crop yield, drought resistance, and nutritional content. By 2014's close, around 1000 research articles had mentioned CRISPR. The technique was used to deactivate genes in human cell lines and cells, explore Candida albicans, modify yeast strains for biofuel production, and genetically alter crops. Researchers assert that genetic sequence manipulation enables reverse engineering, which can positively impact biofuel generation. Additionally, CRISPR can be applied to alter mosquitoes, rendering them incapable of spreading diseases like malaria. Recently, CRISPR-based techniques involving Cas12a have been used to successfully modify a wide range of plant species.
In July 2019, a 34-year-old woman with sickle cell disease became the first patient to be treated with CRISPR for a genetic disorder. In February 2020, advances in HIV treatment were made, with 60-80% of the integrated viral DNA eliminated in mice, and some mice becoming entirely virus-free after LASER ART, a novel anti-retroviral therapy, and CRISPR were used. In March 2020, a patient's eye was injected with a CRISPR-modified virus in an attempt to address Leber congenital amaurosis.
In the future, CRISPR gene editing may have the potential to create new species or resurrect extinct ones from closely related organisms. CRISPR-based re-assessments of gene-disease relationship claims have led to the identification of potentially significant discrepancies. In July 2021, CRISPR gene editing of hiPSCs was employed to investigate the role of MBNL proteins related to DM1.
CRISPR-associated nucleases have proven valuable for molecular testing due to their capacity to target specific nucleic acid sequences amidst a high background of non-target sequences. In 2016, Cas9 nuclease was utilized to eliminate undesired nucleotide sequences in next-generation sequencing libraries, requiring a mere 250 picograms of initial RNA input. Starting in 2017, CRISPR-associated nucleases were employed for direct diagnostic testing of nucleic acids, with sensitivity down to single molecules. CRISPR diversity serves as an analytical target to determine phylogeny and diversity in bacteria, such as xanthomonads. Early detection of plant pathogens through molecular typing of the pathogen's CRISPRs can benefit agriculture.
By linking CRISPR-based diagnostics with additional enzymatic processes, detecting molecules beyond nucleic acids becomes possible. An example of such a coupled technology is SHERLOCK-based Profiling of IN vitro Transcription (SPRINT). SPRINT can be employed to identify various substances, including metabolites in patient samples or contaminants in environmental samples, using either high-throughput methods or portable point-of-care devices. CRISPR/Cas platforms are also being investigated for the detection and inactivation of SARS-CoV-2, the virus responsible for COVID-19. Two comprehensive diagnostic tests, AIOD-CRISPR and the SHERLOCK test, have been developed for SARS-CoV-2. The SHERLOCK test relies on a fluorescently labeled reporter RNA capable of detecting 10 copies per microliter. AIOD-CRISPR assists in robust and highly sensitive visual detection of viral nucleic acid.
CRISPR technology has demonstrated immense potential in various applications, from agriculture and gene editing for medical treatments to molecular diagnostics and pathogen detection. As researchers continue to explore and refine this revolutionary tool, it is expected to play a pivotal role in addressing a wide array of global challenges. As we move forward, the responsible development and use of CRISPR will be essential to harness its full potential and ensure its benefits are accessible to all.
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