The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V ...The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V CRISPR-Cpfl （also known as Cas12a） system is a two-component transcriptional unit （TCTU） in which the Cas9 or Cpf1 protein is expressed from an RNA polymerase （pol） II promoter, whereas the single guide RNA （sgRNA） is typically expressed from a Pol III promoter, such as U6 or U3 promoter.展开更多
Ever since its mechanism was discovered back in 2012,the CRISPR/Cas9 system have revolutionized the field of genome editing.While at first it was seen as a therapeutic tool mostly relevant for curing genetic diseases,...Ever since its mechanism was discovered back in 2012,the CRISPR/Cas9 system have revolutionized the field of genome editing.While at first it was seen as a therapeutic tool mostly relevant for curing genetic diseases,it has been recently shown to also hold the potential to become a clinically relevant therapy for cancer.However,there are multiple challenges that must be addressed prior to clinical testing.Predominantly,the safety of the system when used for in-vivo therapies,including off-target activity and the effects of the double strand break induction on genomic stability.Here,we will focus on the inherent challenges in the CRISPR/Cas9 system and discuss various opportunities to overcoming these challenges.展开更多
文摘The class 2 clustered regularly interspaced short palindromic repeat （CRISPR） systems have been widely used for simultaneous modification of multiple loci in plants. Traditionally, the type II CRISPR-Cas9 or type V CRISPR-Cpfl （also known as Cas12a） system is a two-component transcriptional unit （TCTU） in which the Cas9 or Cpf1 protein is expressed from an RNA polymerase （pol） II promoter, whereas the single guide RNA （sgRNA） is typically expressed from a Pol III promoter, such as U6 or U3 promoter.
文摘Ever since its mechanism was discovered back in 2012,the CRISPR/Cas9 system have revolutionized the field of genome editing.While at first it was seen as a therapeutic tool mostly relevant for curing genetic diseases,it has been recently shown to also hold the potential to become a clinically relevant therapy for cancer.However,there are multiple challenges that must be addressed prior to clinical testing.Predominantly,the safety of the system when used for in-vivo therapies,including off-target activity and the effects of the double strand break induction on genomic stability.Here,we will focus on the inherent challenges in the CRISPR/Cas9 system and discuss various opportunities to overcoming these challenges.
