From Sanger sequencing to high-throughput sequencing, the rapid development of sequencing technology has been providing better technical support for combating crime through forensic DNA analysis. In recent years, the third-generation sequencing technology, mainly based on nanopore sequencing technology, has been widely applied in life science research, in vitro diagnosis, public health, food safety and other fields. Nanopore sequencing technology with super-long reading and real-time sequencing has great potential in the field of forensic genetics. Many authorities and experts have already realized the great potential of nanopore sequencing applications for forensic purposes, although its application in forensic science is still in its infancy. There is little relevant research literature in the field of forensic science, and we should research and explore it further. In this article, the authors attempt to describe the basic principle and characteristics of nanopore sequencing technology, and share the updates of nanopore sequencing-based STR typing, MH typing, mtDNA, DNA methylation and RNA sequencing during the past several years. Meanwhile, non-human genetic material can provide medical examiners with special evidence and clues. The past decade has witnessed the enormous potential of nanopore sequencing technology in non-human forensic genetics. Especially in the areas of microorganisms, plant, and animal forensics, the application of nanopore sequencing to species identification can exert a huge implication, and provide the vital evidence and clues for the public security. In addition, nanopore sequencing has been used to detect viruses at the scene. In the field of forensic genetics, the nanopore sequencing with portability and real-time sequencing makes it most likely to sequence directly of biological samples at the crime scene. This development of the nanopore sequencing has opened up new possibilities by bringing “the laboratory into the field”. This draw the incomparable attraction to the practical application in public security. Moreover, several problems with nanopore sequencing in forensic genetics are discussed, including complex data analysis, high error rate, high sample quality requirements, and analytical methods, and there is a certain distance from the daily application of forensic genetics, which need in-depth research. Finally, we hope that this review can provide a reference for related research and applications, opening up ideas for relevant personnel.