مجله دانشکده پزشکی اصفهان
پیاپی 612 (هفته چهارم فروردین 1400)

Clustered regularly interspaced short palindromic repeats/ CRISPR-associated protein 9 (CRISPR/Cas9)-mediated gene knockout of primary T cell has several limitations for clinical applications. Direct delivery of recombinant Cas9 protein and synthetic gRNA, as a pre-assembled ribonucleoprotein (RNP) complex, has become a potent approach to introduce highly efficient gene editing in primary T cells. In this study, we employed Cas9 RNP-based delivery system for targeted T Cell receptor alpha constant (TRAC) and β2 microglobulin (B2M) genes knockout in human primary T cells.

Specific gRNAs were designed to target the first exons of TRAC and B2M genes. Cas9 protein and respective synthetic gRNAs were then mixed separately, and electroporated into human primary T cells isolated from peripheral blood mononuclear cells (PBMCs). The gene editing efficiency was quantified using tracking of indels by decomposition (TIDE) analysis and flow cytometry.

Three days after electroporation of primary T cells with the TRAC and B2M targeting RNP complexes, TIDE analysis revealed the knockout efficiency of 13-60 percent for the TRAC-targeting gRNAs and 21-53 percent for B2M-targeting gRNAs. Flow cytometry analysis confirmed ~76% and ~27% complete loss of expression for the most efficient gRNAs targeting TRAC (TRAC-gRNA3) and B2M (B2M-gRNA2), respectively.

Our results demonstrate that Cas9 RNP system can be efficiently delivered into primary T cells and result in targeted gene knockout. The protocol described here enables a streamlined and highly efficient solution for maximizing editing efficiency in primary T cells, and simplifies the gene editing process for next-generation immunotherapies

Coronavirus disease 2019 (COVID-19) is the largest and most severe epidemic since the 1918 flu, and its emergence has created unprecedented challenges to public health. Clinical manifestations in patients are very diverse, and can range from asymptomatic disease to severe illness with death. The main purpose of this study was to compare the outcomes of patients with Covid-19 with early pulmonary, gastrointestinal, and neurological symptoms in patients admitted to hospitals in Qazvin City, Iran.

This was a retrospective study with 300 samples, performed after obtaining ethics license and obtaining patient consent. The samples were selected using purposive method. Confirmation of COVID-19 was done due to having a lung scan confirmed by an internal medicine or infectious disease specialist, as well as having a positive polymerase chain reaction (PCR) test. Exclusion criteria also included pregnancy.

The group of patients presented with early pulmonary symptoms were significantly different from the other two groups in terms of mortality, intensive care unit, intubation and hospitalization (P < 0.050).

Patients presenting with early pulmonary symptoms should be given priority in hospitalization.

The terms portfolio and logbook have been widely used in the field of medical education. However, the differences and similarities between them are still under question. In this letter to the editor, an attempt has been made to make the necessary clarification in this regard by reviewing the relevant sources.