دانلود رایگان مقاله انگلیسی ویرایش CRISPR در تحقیقات بیولوژیکی و زیست پزشکی - وایلی 2017

AbstractRecently, clustered regularly interspaced short palindromic repeats (CRISPR) based genomic editing technologies have armed researchers with powerful new tools to biological and biomedical investigations. To further improve and expand its functionality, natural and engineered CRISPR associated 9 proteins (Cas9s) have been investigated, various CRISPR delivery strategies have been tested and optimized, and multiple schemes have been developed to ensure precise mammalian genome editing. Benefiting from those in-depth understanding and further development of CRISPR, versatile CRISPR-based platforms for genome editing have been rapidly developed to advance investigations in biology and biomedicine. In biological research area, CRISPR has been widely adopted in both fundamental and applied research fields, such as accurate base editing, transcriptional regulation and genome-wide screening. In biomedical research area, CRISPR has also shown its extensive applicability in the establishment of animal models for genetic disorders especially those large animals and non-human primates models, and gene therapy to combat virus infectious diseases, to correct monogenic disorders in vivo or in pluripotent cells. In this prospect article, after highlighting recent developments of CRISPR systems, we outline different applications and current limitations of CRISPR use in biological and biomedical investigation. Finally, we provide a perspective for future development and potential risks of this multifunctional technology. This article is protected by copyright.

5. Conclusion

The discovery and characterization of CRISPR systems have transformed the studying in life sciences. The development of new CRISPR associated engineer technologies such as investigating new natural CRISPR enzymes, expanding the targeting scope of Cas9 and improving the DNA specificity of CRISPR-based agents have driven and accelerated this transformation. During the preparing of this manuscript, one group describes a CRISPR/Cas9-based approach for inserting a poly(A) transcriptional terminator into both alleles of a targeted gene to silence protein-coding and non-protein-coding genes, which often play key roles in gene regulation but are difficult to silence via insertion or deletion of short DNA fragments (Liu et al., 2017b). The other group develops CRISPR base editors to knock out genes by changing single nucleotides to create stop codons. CRISPR-STOP-mediated targeted screening demonstrates comparable efficiency to WT Cas9, which indicates the suitability of our approach for genome-wide functional screenings (Kuscu et al., 2017). Recently, efficient, footprint-free conditional genome editing was achieve by encapsulating Cas9 gene on a piggyBac transposon (Wang et al., 2017). Additionally, CRISPR/Cas9 has been used human in preimplantation embryos to edit human genetic disease associated gene- the endogenous β-globin gene (HBB), but off-targets and mosaic by-product were all found in those edited embryos, which highlight the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing (Liang et al., 2015).